Protective window for a confocal microscope?

[Tomek Węgierski]

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Dear All,
I am considering a purchase of a protective window for our Zeiss LSM800 and I would like to hear your advice whether this is a good idea, and if yes, which one to buy.
The reason is our LSM800 is on an inverted stand and we already got a leakage of oil once which went down onto Optovar. And this was despite my regular checks of objectives (both sides) for leaking oil and extensive education of users on how important it is not to use too much oil and to clean lenses, especially on inverted stands.
Zeiss Observer has a dummy slider with an opening for 32 mm filters. I hope, a protective window there should stop the oil, if the leakage happens again.
1) do you think placing a protective window in a confocal microscope is a good idea or not really, because the imaging will be affected? This would be in infinity space, so I guess it should not be detrimental....
2) which characteristics of protective window I should take into consideration (substrate, thickness, coating, flatness, surface quality, parallelism)? Currently I am inclined into protective glasses by Edmund Optics such as 1-mm thick MgF2-coated Sapphire window, which has high transmission, or 2-mm thick MgF2-coated B270 window, which has a bit worse transmission but better surface parameters (and is much cheaper).
best regards,
Tomasz


Tomasz Wegierski, PhD
International Institute of Molecular and Cell Biology
Trojdena 4, 02-109 Warsaw, POLAND
tel: [phone]
fax: [phone]
http://www.iimcb.gov.pl/

[Douglas Richardson]

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I would highly recommend the aquastop system from Zeiss. It is overpriced for some pieces of plastic, but it forms a very tight seal and pays for itself many times over. It is far superior to hair elastics/glove fingers/condoms which we used in the past.

Doug

Get Outlook for Android

________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Tomek Węgierski <[hidden email]>
Sent: Tuesday, October 27, 2020 7:12:10 AM
To: [hidden email] <[hidden email]>
Subject: Protective window for a confocal microscope?

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Dear All,
I am considering a purchase of a protective window for our Zeiss LSM800 and I would like to hear your advice whether this is a good idea, and if yes, which one to buy.
The reason is our LSM800 is on an inverted stand and we already got a leakage of oil once which went down onto Optovar. And this was despite my regular checks of objectives (both sides) for leaking oil and extensive education of users on how important it is not to use too much oil and to clean lenses, especially on inverted stands.
Zeiss Observer has a dummy slider with an opening for 32 mm filters. I hope, a protective window there should stop the oil, if the leakage happens again.
1) do you think placing a protective window in a confocal microscope is a good idea or not really, because the imaging will be affected? This would be in infinity space, so I guess it should not be detrimental....
2) which characteristics of protective window I should take into consideration (substrate, thickness, coating, flatness, surface quality, parallelism)? Currently I am inclined into protective glasses by Edmund Optics such as 1-mm thick MgF2-coated Sapphire window, which has high transmission, or 2-mm thick MgF2-coated B270 window, which has a bit worse transmission but better surface parameters (and is much cheaper).
best regards,
Tomasz


Tomasz Wegierski, PhD
International Institute of Molecular and Cell Biology
Trojdena 4, 02-109 Warsaw, POLAND
tel: [phone]
fax: [phone]
http://www.iimcb.gov.pl/

[Avi Jacob]

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Maybe I misunderstand the issue, but if it's because of oil leaking off oil
objectives and getting into the insides, I use these ponytail elastic
fluffy things on every oil objective. (I don't understand why Doug does not
like em. Doug, maybe you did not use ones that can soak up oil well
enough?).
Just stretch one a bit, and place it on the objective. It will soak up any
oil and when saturated, just throw it out.  Every month or so I switch them
out and I sleep soundly - I have zero oil spilling off the objectives. The
amazon link is just to see specifically what I mean, you can pick these up
all over (took me some minutes to find that exact type). I luckily found a
package with perfectly sized ones (in my daughter's drawer).

https://www.amazon.com/Elastic-Bands-Rubber-Ponytail-Holders/dp/B075FLPVYT
--
Avi Jacob, Ph.D.
Head of The Kanbar Light Microscopy Unit
The Goodman Faculty of Life Sciences
Bar-Ilan University, Ramat-Gan 5290002, Israel




On Tue, Oct 27, 2020 at 1:45 PM Doug Richardson <[hidden email]>
wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> I would highly recommend the aquastop system from Zeiss. It is overpriced
> for some pieces of plastic, but it forms a very tight seal and pays for
> itself many times over. It is far superior to hair elastics/glove
> fingers/condoms which we used in the past.
>
> Doug
>
> Get Outlook for Android
>
> ________________________________
> From: Confocal Microscopy List <[hidden email]> on
> behalf of Tomek Węgierski <[hidden email]>
> Sent: Tuesday, October 27, 2020 7:12:10 AM
> To: [hidden email] <[hidden email]>
> Subject: Protective window for a confocal microscope?
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear All,
> I am considering a purchase of a protective window for our Zeiss LSM800
> and I would like to hear your advice whether this is a good idea, and if
> yes, which one to buy.
> The reason is our LSM800 is on an inverted stand and we already got a
> leakage of oil once which went down onto Optovar. And this was despite my
> regular checks of objectives (both sides) for leaking oil and extensive
> education of users on how important it is not to use too much oil and to
> clean lenses, especially on inverted stands.
> Zeiss Observer has a dummy slider with an opening for 32 mm filters. I
> hope, a protective window there should stop the oil, if the leakage happens
> again.
> 1) do you think placing a protective window in a confocal microscope is a
> good idea or not really, because the imaging will be affected? This would
> be in infinity space, so I guess it should not be detrimental....
> 2) which characteristics of protective window I should take into
> consideration (substrate, thickness, coating, flatness, surface quality,
> parallelism)? Currently I am inclined into protective glasses by Edmund
> Optics such as 1-mm thick MgF2-coated Sapphire window, which has high
> transmission, or 2-mm thick MgF2-coated B270 window, which has a bit worse
> transmission but better surface parameters (and is much cheaper).
> best regards,
> Tomasz
>
>
> Tomasz Wegierski, PhD
> International Institute of Molecular and Cell Biology
> Trojdena 4, 02-109 Warsaw, POLAND
> tel: [phone]
> fax: [phone]
> http://www.iimcb.gov.pl/
>

[Cammer, Michael-2]

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*****

Aquastop works well for big spills when installed correctly and has proven effective on some of our Zeiss scopes.  But we did have a user who spilled over or around it on our Zeiss 710, so it is not foolproof.


Hair elastics are great for the oil.


Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory

NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York, NY  10016

[hidden email]<mailto:[hidden email]>  http://nyulmc.org/micros  http://microscopynotes.com/

Voice direct only, no text or messages:  1-914-309-3270 and 1-646-501-0567



________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Doug Richardson <[hidden email]>
Sent: Tuesday, October 27, 2020 7:41:45 AM
To: [hidden email]
Subject: Re: Protective window for a confocal microscope?

[EXTERNAL]

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*****

I would highly recommend the aquastop system from Zeiss. It is overpriced for some pieces of plastic, but it forms a very tight seal and pays for itself many times over. It is far superior to hair elastics/glove fingers/condoms which we used in the past.

Doug

Get Outlook for Android

________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Tomek Węgierski <[hidden email]>
Sent: Tuesday, October 27, 2020 7:12:10 AM
To: [hidden email] <[hidden email]>
Subject: Protective window for a confocal microscope?

*****
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Post images on https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=o9KAzzxXBHkEPM_YiZYo9mb7m2x6GjWRoUHh-BzcNSg&e=  and include the link in your posting.
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Dear All,
I am considering a purchase of a protective window for our Zeiss LSM800 and I would like to hear your advice whether this is a good idea, and if yes, which one to buy.
The reason is our LSM800 is on an inverted stand and we already got a leakage of oil once which went down onto Optovar. And this was despite my regular checks of objectives (both sides) for leaking oil and extensive education of users on how important it is not to use too much oil and to clean lenses, especially on inverted stands.
Zeiss Observer has a dummy slider with an opening for 32 mm filters. I hope, a protective window there should stop the oil, if the leakage happens again.
1) do you think placing a protective window in a confocal microscope is a good idea or not really, because the imaging will be affected? This would be in infinity space, so I guess it should not be detrimental....
2) which characteristics of protective window I should take into consideration (substrate, thickness, coating, flatness, surface quality, parallelism)? Currently I am inclined into protective glasses by Edmund Optics such as 1-mm thick MgF2-coated Sapphire window, which has high transmission, or 2-mm thick MgF2-coated B270 window, which has a bit worse transmission but better surface parameters (and is much cheaper).
best regards,
Tomasz


Tomasz Wegierski, PhD
International Institute of Molecular and Cell Biology
Trojdena 4, 02-109 Warsaw, POLAND
tel: [phone]
fax: [phone]
https://urldefense.proofpoint.com/v2/url?u=http-3A__www.iimcb.gov.pl_&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=u7qHptX43byvVfMe-Zn04ev5KF4-ToW1ss01pq0eVtM&e=

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[Lemasters, John J.]

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Women in the US at least will recognize "ponytail elastic fluffy things" as "scrunchies", which you can buy under this name from Amazon and other suppliers. We use them also. They are helpful in a multi-user environment where inexperienced users can get overly exuberant in oiling objectives. They should be replaced when oil drips on them.

Best, John

--
John J. Lemasters, MD, PhD
Professor and GlaxoSmithKline Distinguished Endowed Chair
Director, Center for Cell Death, Injury & Regeneration
Departments of Drug Discovery & Biomedical Sciences and Biochemistry & Molecular Biology
Medical University of South Carolina
DD504 Drug Discovery Building
70 President Street, MSC 139
Charleston, SC 29425

Office: 843-876-2360
Lab: 843-876-2354
Fax: [phone]
Email: [hidden email]
https://education.musc.edu/MUSCApps/FacultyDirectory/Lemasters-John

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Avi Jacob
Sent: Tuesday, October 27, 2020 9:06 AM
To: [hidden email]
Subject: Re: Protective window for a confocal microscope?

CAUTION: External

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Post images on https://urldefense.com/v3/__http://www.imgur.com__;!!Ab1_Rw!Skra7YcxZVCzHTYZgYmYyd8oaFrWhQjgg2qsfZlzOaztVPV5nx3K6ZEjzY16Kn-lbJQ$  and include the link in your posting.
*****

Maybe I misunderstand the issue, but if it's because of oil leaking off oil objectives and getting into the insides, I use these ponytail elastic fluffy things on every oil objective. (I don't understand why Doug does not like em. Doug, maybe you did not use ones that can soak up oil well enough?).
Just stretch one a bit, and place it on the objective. It will soak up any oil and when saturated, just throw it out.  Every month or so I switch them out and I sleep soundly - I have zero oil spilling off the objectives. The amazon link is just to see specifically what I mean, you can pick these up all over (took me some minutes to find that exact type). I luckily found a package with perfectly sized ones (in my daughter's drawer).

https://urldefense.com/v3/__https://www.amazon.com/Elastic-Bands-Rubber-Ponytail-Holders/dp/B075FLPVYT__;!!Ab1_Rw!Skra7YcxZVCzHTYZgYmYyd8oaFrWhQjgg2qsfZlzOaztVPV5nx3K6ZEjzY16r31RiXc$
--
Avi Jacob, Ph.D.
Head of The Kanbar Light Microscopy Unit The Goodman Faculty of Life Sciences Bar-Ilan University, Ramat-Gan 5290002, Israel




On Tue, Oct 27, 2020 at 1:45 PM Doug Richardson <[hidden email]>
wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> https://urldefense.com/v3/__http://lists.umn.edu/cgi-bin/wa?A0=confoca
> lmicroscopy__;!!Ab1_Rw!Skra7YcxZVCzHTYZgYmYyd8oaFrWhQjgg2qsfZlzOaztVPV
> 5nx3K6ZEjzY16IDn8MpE$ Post images on
> https://urldefense.com/v3/__http://www.imgur.com__;!!Ab1_Rw!Skra7YcxZVCzHTYZgYmYyd8oaFrWhQjgg2qsfZlzOaztVPV5nx3K6ZEjzY16Kn-lbJQ$  and include the link in your posting.
> *****
>
> I would highly recommend the aquastop system from Zeiss. It is
> overpriced for some pieces of plastic, but it forms a very tight seal
> and pays for itself many times over. It is far superior to hair
> elastics/glove fingers/condoms which we used in the past.
>
> Doug
>
> Get Outlook for Android
>
> ________________________________
> From: Confocal Microscopy List <[hidden email]> on
> behalf of Tomek Węgierski <[hidden email]>
> Sent: Tuesday, October 27, 2020 7:12:10 AM
> To: [hidden email]
> <[hidden email]>
> Subject: Protective window for a confocal microscope?
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> https://urldefense.com/v3/__http://lists.umn.edu/cgi-bin/wa?A0=confoca
> lmicroscopy__;!!Ab1_Rw!Skra7YcxZVCzHTYZgYmYyd8oaFrWhQjgg2qsfZlzOaztVPV
> 5nx3K6ZEjzY16IDn8MpE$ Post images on
> https://urldefense.com/v3/__http://www.imgur.com__;!!Ab1_Rw!Skra7YcxZVCzHTYZgYmYyd8oaFrWhQjgg2qsfZlzOaztVPV5nx3K6ZEjzY16Kn-lbJQ$  and include the link in your posting.
> *****
>
> Dear All,
> I am considering a purchase of a protective window for our Zeiss
> LSM800 and I would like to hear your advice whether this is a good
> idea, and if yes, which one to buy.
> The reason is our LSM800 is on an inverted stand and we already got a
> leakage of oil once which went down onto Optovar. And this was despite
> my regular checks of objectives (both sides) for leaking oil and
> extensive education of users on how important it is not to use too
> much oil and to clean lenses, especially on inverted stands.
> Zeiss Observer has a dummy slider with an opening for 32 mm filters. I
> hope, a protective window there should stop the oil, if the leakage
> happens again.
> 1) do you think placing a protective window in a confocal microscope
> is a good idea or not really, because the imaging will be affected?
> This would be in infinity space, so I guess it should not be detrimental....
> 2) which characteristics of protective window I should take into
> consideration (substrate, thickness, coating, flatness, surface
> quality, parallelism)? Currently I am inclined into protective glasses
> by Edmund Optics such as 1-mm thick MgF2-coated Sapphire window, which
> has high transmission, or 2-mm thick MgF2-coated B270 window, which
> has a bit worse transmission but better surface parameters (and is much cheaper).
> best regards,
> Tomasz
>
>
> Tomasz Wegierski, PhD
> International Institute of Molecular and Cell Biology Trojdena 4,
> 02-109 Warsaw, POLAND
> tel: [phone]
> fax: [phone]
> https://urldefense.com/v3/__http://www.iimcb.gov.pl/__;!!Ab1_Rw!Skra7Y
> cxZVCzHTYZgYmYyd8oaFrWhQjgg2qsfZlzOaztVPV5nx3K6ZEjzY16JqeOqTQ$
>

[Gary Laevsky]

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The hair ties work great with objectives that have enough of a neck for the
tie to sit on, next to the barrel.

Our 40X objective (most frequently used) has no/very small neck, so the
hair ties don't work.

I fairly regularly (bi-weekly) have to take the objective off, store it in
it's case upside down, and put it in a 37C oven.  Ridiculous, in a bad way,
how much oil comes out.  The 40X also doesn't have "lip" that acts as a bit
of a reservoir (unlike the 60 and 100).

On one of my heavily used systems, I have to go so far as to dismantle the
top (stage, objective turret, and fluorescent turret) to get to the piece
of glass that prevents leakage into the body (amazingly awesome feature of
the stand).  Takes me 15 minutes.  Although I shouldn't have to do that ...

On Tue, Oct 27, 2020 at 9:13 AM Cammer, Michael <
[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Aquastop works well for big spills when installed correctly and has proven
> effective on some of our Zeiss scopes.  But we did have a user who spilled
> over or around it on our Zeiss 710, so it is not foolproof.
>
>
> Hair elastics are great for the oil.
>
>
> Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory
>
> NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York, NY
> 10016
>
> [hidden email]<mailto:[hidden email]>
> http://nyulmc.org/micros  http://microscopynotes.com/
>
> Voice direct only, no text or messages:  1-914-309-3270 and 1-646-501-0567
>
>
>
> ________________________________
> From: Confocal Microscopy List <[hidden email]> on
> behalf of Doug Richardson <[hidden email]>
> Sent: Tuesday, October 27, 2020 7:41:45 AM
> To: [hidden email]
> Subject: Re: Protective window for a confocal microscope?
>
> [EXTERNAL]
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
>
> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=k-eCFZaqV2J3Nbh8c16znCG7uPPgwni6bBeSRyqYRAE&e=
> Post images on
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=o9KAzzxXBHkEPM_YiZYo9mb7m2x6GjWRoUHh-BzcNSg&e=
> and include the link in your posting.
> *****
>
> I would highly recommend the aquastop system from Zeiss. It is overpriced
> for some pieces of plastic, but it forms a very tight seal and pays for
> itself many times over. It is far superior to hair elastics/glove
> fingers/condoms which we used in the past.
>
> Doug
>
> Get Outlook for Android
>
> ________________________________
> From: Confocal Microscopy List <[hidden email]> on
> behalf of Tomek Węgierski <[hidden email]>
> Sent: Tuesday, October 27, 2020 7:12:10 AM
> To: [hidden email] <[hidden email]>
> Subject: Protective window for a confocal microscope?
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
>
> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=k-eCFZaqV2J3Nbh8c16znCG7uPPgwni6bBeSRyqYRAE&e=
> Post images on
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=o9KAzzxXBHkEPM_YiZYo9mb7m2x6GjWRoUHh-BzcNSg&e=
> and include the link in your posting.
> *****
>
> Dear All,
> I am considering a purchase of a protective window for our Zeiss LSM800
> and I would like to hear your advice whether this is a good idea, and if
> yes, which one to buy.
> The reason is our LSM800 is on an inverted stand and we already got a
> leakage of oil once which went down onto Optovar. And this was despite my
> regular checks of objectives (both sides) for leaking oil and extensive
> education of users on how important it is not to use too much oil and to
> clean lenses, especially on inverted stands.
> Zeiss Observer has a dummy slider with an opening for 32 mm filters. I
> hope, a protective window there should stop the oil, if the leakage happens
> again.
> 1) do you think placing a protective window in a confocal microscope is a
> good idea or not really, because the imaging will be affected? This would
> be in infinity space, so I guess it should not be detrimental....
> 2) which characteristics of protective window I should take into
> consideration (substrate, thickness, coating, flatness, surface quality,
> parallelism)? Currently I am inclined into protective glasses by Edmund
> Optics such as 1-mm thick MgF2-coated Sapphire window, which has high
> transmission, or 2-mm thick MgF2-coated B270 window, which has a bit worse
> transmission but better surface parameters (and is much cheaper).
> best regards,
> Tomasz
>
>
> Tomasz Wegierski, PhD
> International Institute of Molecular and Cell Biology
> Trojdena 4, 02-109 Warsaw, POLAND
> tel: [phone]
> fax: [phone]
>
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.iimcb.gov.pl_&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=u7qHptX43byvVfMe-Zn04ev5KF4-ToW1ss01pq0eVtM&e=
>
> ------------------------------------------------------------
> This email message, including any attachments, is for the sole use of the
> intended recipient(s) and may contain information that is proprietary,
> confidential, and exempt from disclosure under applicable law. Any
> unauthorized review, use, disclosure, or distribution is prohibited. If you
> have received this email in error please notify the sender by return email
> and delete the original message. Please note, the recipient should check
> this email and any attachments for the presence of viruses. The
> organization accepts no liability for any damage caused by any virus
> transmitted by this email.
> =================================
>

--
Best,

Gary Laevsky, Ph.D.
Director, Confocal Imaging Facility
Nikon Center of Excellence
Co-Founder, North Atlantic Microscopy Society (NAMS)
https://namsmicroscopy.com/
Dept. of Molecular Biology
Washington Rd.
Princeton University
Princeton, New Jersey, 08544-1014
(O) 609 258 5432
(C) 508 507 1310

[Michael Stanley]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

in the 'old days' in the core we used to cut filter paper and then bore a
hole with a cork-bore slightly smaller than the lens.  slide 2-3 (or as many
as will fit) layers of the filter paper over the objective.  this will not
stop a "flood" of oil (which sounds very possible for some) but it will hold
much more than you would think.  and, the most important feature of this
little trick is that the oil will show very quickly on the paper and
hopefully will trigger a response.!

some of the darker filter papers will show the oil better, but this will
vary by manufacturer.

no commercial interest here, just trying to protect all the optics in the
system.

michael

c. michael stanley, phd
senior applications scientist
chroma technology corp.
10 imtec lane
bellows falls,  vt    05101
[hidden email]
[hidden email]


-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf
Of Gary Laevsky
Sent: Tuesday, October 27, 2020 10:19 AM
To: [hidden email]
Subject: Re: Protective window for a confocal microscope?

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

The hair ties work great with objectives that have enough of a neck for the
tie to sit on, next to the barrel.

Our 40X objective (most frequently used) has no/very small neck, so the hair
ties don't work.

I fairly regularly (bi-weekly) have to take the objective off, store it in
it's case upside down, and put it in a 37C oven.  Ridiculous, in a bad way,
how much oil comes out.  The 40X also doesn't have "lip" that acts as a bit
of a reservoir (unlike the 60 and 100).

On one of my heavily used systems, I have to go so far as to dismantle the
top (stage, objective turret, and fluorescent turret) to get to the piece of
glass that prevents leakage into the body (amazingly awesome feature of the
stand).  Takes me 15 minutes.  Although I shouldn't have to do that ...

On Tue, Oct 27, 2020 at 9:13 AM Cammer, Michael <
[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Aquastop works well for big spills when installed correctly and has
> proven effective on some of our Zeiss scopes.  But we did have a user
> who spilled over or around it on our Zeiss 710, so it is not foolproof.
>
>
> Hair elastics are great for the oil.
>
>
> Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory
>
> NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York,
> NY
> 10016
>
> [hidden email]<mailto:[hidden email]>
> http://nyulmc.org/micros  http://microscopynotes.com/
>
> Voice direct only, no text or messages:  1-914-309-3270 and
> 1-646-501-0567
>
>
>
> ________________________________
> From: Confocal Microscopy List <[hidden email]> on
> behalf of Doug Richardson <[hidden email]>
> Sent: Tuesday, October 27, 2020 7:41:45 AM
> To: [hidden email]
> Subject: Re: Protective window for a confocal microscope?
>
> [EXTERNAL]
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
>
> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-
> 2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeEl
> Zfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=ke
> g1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=k-eCFZaqV2J3Nbh8c16znCG7uP
> Pgwni6bBeSRyqYRAE&e=
> Post images on
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=Dw
> IFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC5
> 0tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-d
> FQ&s=o9KAzzxXBHkEPM_YiZYo9mb7m2x6GjWRoUHh-BzcNSg&e=
> and include the link in your posting.
> *****
>
> I would highly recommend the aquastop system from Zeiss. It is
> overpriced for some pieces of plastic, but it forms a very tight seal
> and pays for itself many times over. It is far superior to hair
> elastics/glove fingers/condoms which we used in the past.
>
> Doug
>
> Get Outlook for Android
>
> ________________________________
> From: Confocal Microscopy List <[hidden email]> on
> behalf of Tomek Węgierski <[hidden email]>
> Sent: Tuesday, October 27, 2020 7:12:10 AM
> To: [hidden email]
> <[hidden email]>
> Subject: Protective window for a confocal microscope?
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
>
> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-
> 2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeEl
> Zfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=ke
> g1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=k-eCFZaqV2J3Nbh8c16znCG7uP
> Pgwni6bBeSRyqYRAE&e=
> Post images on
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=Dw
> IFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC5
> 0tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-d
> FQ&s=o9KAzzxXBHkEPM_YiZYo9mb7m2x6GjWRoUHh-BzcNSg&e=
> and include the link in your posting.
> *****
>
> Dear All,
> I am considering a purchase of a protective window for our Zeiss
> LSM800 and I would like to hear your advice whether this is a good
> idea, and if yes, which one to buy.
> The reason is our LSM800 is on an inverted stand and we already got a
> leakage of oil once which went down onto Optovar. And this was despite
> my regular checks of objectives (both sides) for leaking oil and
> extensive education of users on how important it is not to use too
> much oil and to clean lenses, especially on inverted stands.
> Zeiss Observer has a dummy slider with an opening for 32 mm filters. I
> hope, a protective window there should stop the oil, if the leakage
> happens again.
> 1) do you think placing a protective window in a confocal microscope
> is a good idea or not really, because the imaging will be affected?
> This would be in infinity space, so I guess it should not be
> detrimental....
> 2) which characteristics of protective window I should take into
> consideration (substrate, thickness, coating, flatness, surface
> quality, parallelism)? Currently I am inclined into protective glasses
> by Edmund Optics such as 1-mm thick MgF2-coated Sapphire window, which
> has high transmission, or 2-mm thick MgF2-coated B270 window, which
> has a bit worse transmission but better surface parameters (and is much
> cheaper).
> best regards,
> Tomasz
>
>
> Tomasz Wegierski, PhD
> International Institute of Molecular and Cell Biology Trojdena 4,
> 02-109 Warsaw, POLAND
> tel: [phone]
> fax: [phone]
>
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.iimcb.gov.pl_&
> d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDi
> PlC50tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNB
> tS-dFQ&s=u7qHptX43byvVfMe-Zn04ev5KF4-ToW1ss01pq0eVtM&e=
>
> ------------------------------------------------------------
> This email message, including any attachments, is for the sole use of
> the intended recipient(s) and may contain information that is
> proprietary, confidential, and exempt from disclosure under applicable
> law. Any unauthorized review, use, disclosure, or distribution is
> prohibited. If you have received this email in error please notify the
> sender by return email and delete the original message. Please note,
> the recipient should check this email and any attachments for the
> presence of viruses. The organization accepts no liability for any
> damage caused by any virus transmitted by this email.
> =================================
>

--
Best,

Gary Laevsky, Ph.D.
Director, Confocal Imaging Facility
Nikon Center of Excellence
Co-Founder, North Atlantic Microscopy Society (NAMS)
https://namsmicroscopy.com/ Dept. of Molecular Biology Washington Rd.
Princeton University
Princeton, New Jersey, 08544-1014
(O) 609 258 5432
(C) 508 507 1310

[Vickie Frohlich]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Just my 2 cents.
Scrunchies work well but are not fool-proof.  Cotton not polyester material are more absorbent.
In addition to scrunchies I suggest wrapping the lens barrel near the nose of the objective with plumbers tape to prevent oil from seeping into the barrel.  It can be done so that the lens retraction still functions.  Then once the lens is on the nosepiece of the scope the plumbers tape can be wrapped there as well.

I've also used thin rubber sheets or parafilm sheets, as well as filter paper (thanks Mike S.!), to draw oil away from the lens and nosepiece.

Best method, however, is to teach users proper oil application and ensure they clean up after themselves.

Regards,
Vickie

Victoria Centonze Frohlich, PhD
Director, Light Microscopy Shared Resource
Cell & Tissue Imaging Center
St. Jude Children’s Research Hospital
262 Danny Thomas Place
MS 312
Memphis, TN 38105

Office: D1052D
Phone and Digital Fax: [phone]
Cell: 1-901-907-2596
Paper Fax: [phone]
Email: [hidden email]

CTIC-LM Location: D1055 and D1056
Facility Phone: [phone]

http://home.stjude.org/cell-tissue-imaging/Pages/default.aspx

Facility Acknowledgement for publications:
“Images were acquired at the Cell & Tissue Imaging Center which is supported by SJCRH and NCI P30 CA021765.”

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Michael Stanley
Sent: Tuesday, October 27, 2020 8:19 AM
To: [hidden email]
Subject: Re: Protective window for a confocal microscope?

Caution: External Sender


*****
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Post images on https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=DKQAcpfaPdTDony5B7X%2Fdi%2BCVChjA7voJxvlDKq9suo%3D&amp;reserved=0 and include the link in your posting.
*****

in the 'old days' in the core we used to cut filter paper and then bore a hole with a cork-bore slightly smaller than the lens.  slide 2-3 (or as many as will fit) layers of the filter paper over the objective.  this will not stop a "flood" of oil (which sounds very possible for some) but it will hold much more than you would think.  and, the most important feature of this little trick is that the oil will show very quickly on the paper and hopefully will trigger a response.!

some of the darker filter papers will show the oil better, but this will vary by manufacturer.

no commercial interest here, just trying to protect all the optics in the system.

michael

c. michael stanley, phd
senior applications scientist
chroma technology corp.
10 imtec lane
bellows falls,  vt    05101
[hidden email]
[hidden email]


-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Gary Laevsky
Sent: Tuesday, October 27, 2020 10:19 AM
To: [hidden email]
Subject: Re: Protective window for a confocal microscope?

*****
To join, leave or search the confocal microscopy listserv, go to:
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Post images on https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=yhWCT39l%2FiZJ9i8H0f4EM7MIstxliSvBVsjVQVNN8cI%3D&amp;reserved=0 and include the link in your posting.
*****

The hair ties work great with objectives that have enough of a neck for the tie to sit on, next to the barrel.

Our 40X objective (most frequently used) has no/very small neck, so the hair ties don't work.

I fairly regularly (bi-weekly) have to take the objective off, store it in it's case upside down, and put it in a 37C oven.  Ridiculous, in a bad way, how much oil comes out.  The 40X also doesn't have "lip" that acts as a bit of a reservoir (unlike the 60 and 100).

On one of my heavily used systems, I have to go so far as to dismantle the top (stage, objective turret, and fluorescent turret) to get to the piece of glass that prevents leakage into the body (amazingly awesome feature of the stand).  Takes me 15 minutes.  Although I shouldn't have to do that ...

On Tue, Oct 27, 2020 at 9:13 AM Cammer, Michael < [hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists
> .umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=04%7C01%7Cv
> ictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C223
> 40fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573459000%7CUnknown%7
> CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXV
> CI6Mn0%3D%7C1000&amp;sdata=q2oFBgmsbEfra%2BIBWB6i8xwEJ%2B8SF%2F59WSo3Y
> %2BKvFKg%3D&amp;reserved=0 Post images on
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=yhWCT39l%2FiZJ9i8H0f4EM7MIstxliSvBVsjVQVNN8cI%3D&amp;reserved=0 and include the link in your posting.
> *****
>
> Aquastop works well for big spills when installed correctly and has
> proven effective on some of our Zeiss scopes.  But we did have a user
> who spilled over or around it on our Zeiss 710, so it is not foolproof.
>
>
> Hair elastics are great for the oil.
>
>
> Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory
>
> NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York,
> NY
> 10016
>
> [hidden email]<mailto:[hidden email]>
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fnyulm
> c.org%2Fmicros&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b8
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> C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLC
> JQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=Ud4O3pAe
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> 0%7C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDA
> iLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=e6srs
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>
> Voice direct only, no text or messages:  1-914-309-3270 and
> 1-646-501-0567
>
>
>
> ________________________________
> From: Confocal Microscopy List <[hidden email]> on
> behalf of Doug Richardson <[hidden email]>
> Sent: Tuesday, October 27, 2020 7:41:45 AM
> To: [hidden email]
> Subject: Re: Protective window for a confocal microscope?
>
> [EXTERNAL]
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
>
> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-
> 2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeEl
> Zfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=ke
> g1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=k-eCFZaqV2J3Nbh8c16znCG7uP
> Pgwni6bBeSRyqYRAE&e=
> Post images on
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=Dw
> IFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC5
> 0tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-d
> FQ&s=o9KAzzxXBHkEPM_YiZYo9mb7m2x6GjWRoUHh-BzcNSg&e=
> and include the link in your posting.
> *****
>
> I would highly recommend the aquastop system from Zeiss. It is
> overpriced for some pieces of plastic, but it forms a very tight seal
> and pays for itself many times over. It is far superior to hair
> elastics/glove fingers/condoms which we used in the past.
>
> Doug
>
> Get Outlook for Android
>
> ________________________________
> From: Confocal Microscopy List <[hidden email]> on
> behalf of Tomek Węgierski <[hidden email]>
> Sent: Tuesday, October 27, 2020 7:12:10 AM
> To: [hidden email]
> <[hidden email]>
> Subject: Protective window for a confocal microscope?
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
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>
> Dear All,
> I am considering a purchase of a protective window for our Zeiss
> LSM800 and I would like to hear your advice whether this is a good
> idea, and if yes, which one to buy.
> The reason is our LSM800 is on an inverted stand and we already got a
> leakage of oil once which went down onto Optovar. And this was despite
> my regular checks of objectives (both sides) for leaking oil and
> extensive education of users on how important it is not to use too
> much oil and to clean lenses, especially on inverted stands.
> Zeiss Observer has a dummy slider with an opening for 32 mm filters. I
> hope, a protective window there should stop the oil, if the leakage
> happens again.
> 1) do you think placing a protective window in a confocal microscope
> is a good idea or not really, because the imaging will be affected?
> This would be in infinity space, so I guess it should not be
> detrimental....
> 2) which characteristics of protective window I should take into
> consideration (substrate, thickness, coating, flatness, surface
> quality, parallelism)? Currently I am inclined into protective glasses
> by Edmund Optics such as 1-mm thick MgF2-coated Sapphire window, which
> has high transmission, or 2-mm thick MgF2-coated B270 window, which
> has a bit worse transmission but better surface parameters (and is
> much cheaper).
> best regards,
> Tomasz
>
>
> Tomasz Wegierski, PhD
> International Institute of Molecular and Cell Biology Trojdena 4,
> 02-109 Warsaw, POLAND
> tel: [phone]
> fax: [phone]
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Best,

Gary Laevsky, Ph.D.
Director, Confocal Imaging Facility
Nikon Center of Excellence
Co-Founder, North Atlantic Microscopy Society (NAMS)
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I have to second Doug's recommendation for the Aquastop from Zeiss.  We use this on both of our inverted confocals and I have found it to be the most robust protection I've ever used.  In our core, it has held up to exposure to oil for well over 2 weeks while I was away on vacation and before I got a chance to inspect the system after returning.

The difference between the Aquastop and other solutions such as hair ties or scrunchies is that the latter only really works for small amounts of oil over very short periods of time.  If you have a user that over oils for one imaging session lasting an hour or so - these scrunchies will work as long as you immediately clean up.

They fail to stop oil penetration over long periods of time with multiple users.  Oil that gets down into your optivar is chronic over oiling - a scrunchie is no match for that.  Under normal conditions I regularly inspect the instruments - and even after exhausting levels of training some users just don't get it, are careless or just plain don't care.

So we do our best to regularly inspect - but the Aquastop has prevented more than a few situations where oil could have ruined our objectives.

Good luck!

-Jason

Jason M. Kirk
Technical Director, Optical Imaging & Vital Microscopy Core (OiVM)
Baylor College of Medicine
Ph: 713.798.6486
Email: [hidden email]
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[Gary Laevsky]

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Does this Holy Grail of oil protection fit non-Zeiss stands?

Best,

Gary

> On Oct 27, 2020, at 11:55 AM, Jason M. Kirk <[hidden email]> wrote:
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> I have to second Doug's recommendation for the Aquastop from Zeiss.  We use this on both of our inverted confocals and I have found it to be the most robust protection I've ever used.  In our core, it has held up to exposure to oil for well over 2 weeks while I was away on vacation and before I got a chance to inspect the system after returning.
>
> The difference between the Aquastop and other solutions such as hair ties or scrunchies is that the latter only really works for small amounts of oil over very short periods of time.  If you have a user that over oils for one imaging session lasting an hour or so - these scrunchies will work as long as you immediately clean up.
>
> They fail to stop oil penetration over long periods of time with multiple users.  Oil that gets down into your optivar is chronic over oiling - a scrunchie is no match for that.  Under normal conditions I regularly inspect the instruments - and even after exhausting levels of training some users just don't get it, are careless or just plain don't care.
>
> So we do our best to regularly inspect - but the Aquastop has prevented more than a few situations where oil could have ruined our objectives.
>
> Good luck!
>
> -Jason
>
> Jason M. Kirk
> Technical Director, Optical Imaging & Vital Microscopy Core (OiVM)
> Baylor College of Medicine
> Ph: 713.798.6486
> Email: [hidden email]
> http://www.bcm.edu/oivm

[Tomek Węgierski]

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Dear All,

Thank you very much for so many responses, although the discussion went a bit into a direction that I did not anticipate ??
But yes, it is all about preventing oil from getting into the microscope. Our 40x/1.3 Fluar lens takes a lot of oil and I frequently have to clean its bottom side from oil. In contrast, I never saw any oil on the back of 63x/1.4 even though it is used as frequently as the 40x.

I have to admit I have been skeptical about hair elastics, because when they saturate with oil they will just let it go. IMHO, it is replacing "oil cleaning" approach with "oil buffering" approach. The "cleaning" approach has worked for us for many years. It failed when an unexperienced user (who apparently did not take seriously what I was talking during the training) made two mistakes: did not clean the lens and let it stay in the optic path after the work instead of changing for 10x. In addition, it was a holiday time, the system was not used for the next few days, giving the oil plenty of time. So, it was a coincidence of several factors, but it can happen again.

I think, hearing so many positive opinions about hair elastics, I will give them the 2nd chance in our facility. Aqua-Stop in my opinion is overpriced, and we do not currently do any experiments under perfusion to justify such purchase (this may change in the future). However, I would not like to give up on the protective window strategy. I think placing a glass element in the infinity space is not unusual. It would function as the last stronghold against the oil. And it would be considerably cheaper than Aqua-Stop. I would of course test PSF. If PSF is not affected, I guess the protective window is OK with imaging. So, if you still have some advice which one would be most suitable, I would appreciate it.

Best,
Tomasz

Tomasz Wegierski, PhD
International Institute of Molecular and Cell Biology
Trojdena 4, 02-109 Warsaw, POLAND
tel: [phone]
fax: [phone]
http://www.iimcb.gov.pl/
________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Frohlich, Victoria <[hidden email]>
Sent: Tuesday, October 27, 2020 4:38 PM
To: [hidden email] <[hidden email]>
Subject: Re: Protective window for a confocal microscope?

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Just my 2 cents.
Scrunchies work well but are not fool-proof.  Cotton not polyester material are more absorbent.
In addition to scrunchies I suggest wrapping the lens barrel near the nose of the objective with plumbers tape to prevent oil from seeping into the barrel.  It can be done so that the lens retraction still functions.  Then once the lens is on the nosepiece of the scope the plumbers tape can be wrapped there as well.

I've also used thin rubber sheets or parafilm sheets, as well as filter paper (thanks Mike S.!), to draw oil away from the lens and nosepiece.

Best method, however, is to teach users proper oil application and ensure they clean up after themselves.

Regards,
Vickie

Victoria Centonze Frohlich, PhD
Director, Light Microscopy Shared Resource
Cell & Tissue Imaging Center
St. Jude Children’s Research Hospital
262 Danny Thomas Place
MS 312
Memphis, TN 38105

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“Images were acquired at the Cell & Tissue Imaging Center which is supported by SJCRH and NCI P30 CA021765.”

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Michael Stanley
Sent: Tuesday, October 27, 2020 8:19 AM
To: [hidden email]
Subject: Re: Protective window for a confocal microscope?

Caution: External Sender


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in the 'old days' in the core we used to cut filter paper and then bore a hole with a cork-bore slightly smaller than the lens.  slide 2-3 (or as many as will fit) layers of the filter paper over the objective.  this will not stop a "flood" of oil (which sounds very possible for some) but it will hold much more than you would think.  and, the most important feature of this little trick is that the oil will show very quickly on the paper and hopefully will trigger a response.!

some of the darker filter papers will show the oil better, but this will vary by manufacturer.

no commercial interest here, just trying to protect all the optics in the system.

michael

c. michael stanley, phd
senior applications scientist
chroma technology corp.
10 imtec lane
bellows falls,  vt    05101
[hidden email]
[hidden email]


-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Gary Laevsky
Sent: Tuesday, October 27, 2020 10:19 AM
To: [hidden email]
Subject: Re: Protective window for a confocal microscope?

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The hair ties work great with objectives that have enough of a neck for the tie to sit on, next to the barrel.

Our 40X objective (most frequently used) has no/very small neck, so the hair ties don't work.

I fairly regularly (bi-weekly) have to take the objective off, store it in it's case upside down, and put it in a 37C oven.  Ridiculous, in a bad way, how much oil comes out.  The 40X also doesn't have "lip" that acts as a bit of a reservoir (unlike the 60 and 100).

On one of my heavily used systems, I have to go so far as to dismantle the top (stage, objective turret, and fluorescent turret) to get to the piece of glass that prevents leakage into the body (amazingly awesome feature of the stand).  Takes me 15 minutes.  Although I shouldn't have to do that ...

On Tue, Oct 27, 2020 at 9:13 AM Cammer, Michael < [hidden email]> wrote:

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> *****
>
> Aquastop works well for big spills when installed correctly and has
> proven effective on some of our Zeiss scopes.  But we did have a user
> who spilled over or around it on our Zeiss 710, so it is not foolproof.
>
>
> Hair elastics are great for the oil.
>
>
> Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory
>
> NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York,
> NY
> 10016
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> ________________________________
> From: Confocal Microscopy List <[hidden email]> on
> behalf of Doug Richardson <[hidden email]>
> Sent: Tuesday, October 27, 2020 7:41:45 AM
> To: [hidden email]
> Subject: Re: Protective window for a confocal microscope?
>
> [EXTERNAL]
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
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>
> I would highly recommend the aquastop system from Zeiss. It is
> overpriced for some pieces of plastic, but it forms a very tight seal
> and pays for itself many times over. It is far superior to hair
> elastics/glove fingers/condoms which we used in the past.
>
> Doug
>
> Get Outlook for Android
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> ________________________________
> From: Confocal Microscopy List <[hidden email]> on
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>
> Dear All,
> I am considering a purchase of a protective window for our Zeiss
> LSM800 and I would like to hear your advice whether this is a good
> idea, and if yes, which one to buy.
> The reason is our LSM800 is on an inverted stand and we already got a
> leakage of oil once which went down onto Optovar. And this was despite
> my regular checks of objectives (both sides) for leaking oil and
> extensive education of users on how important it is not to use too
> much oil and to clean lenses, especially on inverted stands.
> Zeiss Observer has a dummy slider with an opening for 32 mm filters. I
> hope, a protective window there should stop the oil, if the leakage
> happens again.
> 1) do you think placing a protective window in a confocal microscope
> is a good idea or not really, because the imaging will be affected?
> This would be in infinity space, so I guess it should not be
> detrimental....
> 2) which characteristics of protective window I should take into
> consideration (substrate, thickness, coating, flatness, surface
> quality, parallelism)? Currently I am inclined into protective glasses
> by Edmund Optics such as 1-mm thick MgF2-coated Sapphire window, which
> has high transmission, or 2-mm thick MgF2-coated B270 window, which
> has a bit worse transmission but better surface parameters (and is
> much cheaper).
> best regards,
> Tomasz
>
>
> Tomasz Wegierski, PhD
> International Institute of Molecular and Cell Biology Trojdena 4,
> 02-109 Warsaw, POLAND
> tel: [phone]
> fax: [phone]
>
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.iimcb.gov.pl_&
> d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDi
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Best,

Gary Laevsky, Ph.D.
Director, Confocal Imaging Facility
Nikon Center of Excellence
Co-Founder, North Atlantic Microscopy Society (NAMS)
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[Zdenek Svindrych-2]

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Hi Tomasz,

back to your question. The filter holder is there to hold a filter. Any
decent glass or fused silica AR window should do the trick.
As far as I remember, the hole is quite big, around 30 mm diameter (bigger
than the standard 25 mm filters), which may limit your options.

Most filters and windows have specified surface flatness (reflected wave
error), but it's actually transmitted wave error (TWE) that matters here.
Usually, windows with single digit flatness (in lambda or fringe units)
will have TWE << 1/5 lambda, and should be OK.

Thicker substrate (2 - 3 mm) usually means precision optics, and is a safe
bet. Hard coating is a must, if you want to wipe oil off of it regularly.
All reputable brands use hard coatings these days, 1% loss (per pass) is
OK.

Sapphire is hard to work with and comes with a price premium. Fused silica,
e.g.
https://www.edmundoptics.com/p/30mm-dia-vis-nir-coated-1lambda-fused-silica-window/10158/
(no commercial interest) looks like a good option, but BK-7 will work well,
too.

If you happen to have a piece of AR-coated glass at hand, just put it there
and look at some beads. If you're not able to tell the difference, you're
good to go. You can even try a glass coverslip, the 4% loss (the excitation
loss is not an issue, you can always crank up the laser) will be hardly
noticeable. Coverslips are not polished, so the TWE is generally bad, but
with some luck you can find a good one in the stack...

Best, zdenek


On Tue, Oct 27, 2020 at 1:18 PM Tomek Węgierski <[hidden email]>
wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear All,
>
> Thank you very much for so many responses, although the discussion went a
> bit into a direction that I did not anticipate ??
> But yes, it is all about preventing oil from getting into the microscope.
> Our 40x/1.3 Fluar lens takes a lot of oil and I frequently have to clean
> its bottom side from oil. In contrast, I never saw any oil on the back of
> 63x/1.4 even though it is used as frequently as the 40x.
>
> I have to admit I have been skeptical about hair elastics, because when
> they saturate with oil they will just let it go. IMHO, it is replacing "oil
> cleaning" approach with "oil buffering" approach. The "cleaning" approach
> has worked for us for many years. It failed when an unexperienced user (who
> apparently did not take seriously what I was talking during the training)
> made two mistakes: did not clean the lens and let it stay in the optic path
> after the work instead of changing for 10x. In addition, it was a holiday
> time, the system was not used for the next few days, giving the oil plenty
> of time. So, it was a coincidence of several factors, but it can happen
> again.
>
> I think, hearing so many positive opinions about hair elastics, I will
> give them the 2nd chance in our facility. Aqua-Stop in my opinion is
> overpriced, and we do not currently do any experiments under perfusion to
> justify such purchase (this may change in the future). However, I would not
> like to give up on the protective window strategy. I think placing a glass
> element in the infinity space is not unusual. It would function as the last
> stronghold against the oil. And it would be considerably cheaper than
> Aqua-Stop. I would of course test PSF. If PSF is not affected, I guess the
> protective window is OK with imaging. So, if you still have some advice
> which one would be most suitable, I would appreciate it.
>
> Best,
> Tomasz
>
> Tomasz Wegierski, PhD
> International Institute of Molecular and Cell Biology
> Trojdena 4, 02-109 Warsaw, POLAND
> tel: [phone]
> fax: [phone]
> http://www.iimcb.gov.pl/
> ________________________________
> From: Confocal Microscopy List <[hidden email]> on
> behalf of Frohlich, Victoria <[hidden email]>
> Sent: Tuesday, October 27, 2020 4:38 PM
> To: [hidden email] <[hidden email]>
> Subject: Re: Protective window for a confocal microscope?
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Just my 2 cents.
> Scrunchies work well but are not fool-proof.  Cotton not polyester
> material are more absorbent.
> In addition to scrunchies I suggest wrapping the lens barrel near the nose
> of the objective with plumbers tape to prevent oil from seeping into the
> barrel.  It can be done so that the lens retraction still functions.  Then
> once the lens is on the nosepiece of the scope the plumbers tape can be
> wrapped there as well.
>
> I've also used thin rubber sheets or parafilm sheets, as well as filter
> paper (thanks Mike S.!), to draw oil away from the lens and nosepiece.
>
> Best method, however, is to teach users proper oil application and ensure
> they clean up after themselves.
>
> Regards,
> Vickie
>
> Victoria Centonze Frohlich, PhD
> Director, Light Microscopy Shared Resource
> Cell & Tissue Imaging Center
> St. Jude Children’s Research Hospital
> 262 Danny Thomas Place
> MS 312
> Memphis, TN 38105
>
> Office: D1052D
> Phone and Digital Fax: [phone]
> Cell: 1-901-907-2596
> Paper Fax: [phone]
> Email: [hidden email]
>
> CTIC-LM Location: D1055 and D1056
> Facility Phone: [phone]
>
> http://home.stjude.org/cell-tissue-imaging/Pages/default.aspx
>
> Facility Acknowledgement for publications:
> “Images were acquired at the Cell & Tissue Imaging Center which is
> supported by SJCRH and NCI P30 CA021765.”
>
> -----Original Message-----
> From: Confocal Microscopy List <[hidden email]> On
> Behalf Of Michael Stanley
> Sent: Tuesday, October 27, 2020 8:19 AM
> To: [hidden email]
> Subject: Re: Protective window for a confocal microscope?
>
> Caution: External Sender
>
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
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> Post images on
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=DKQAcpfaPdTDony5B7X%2Fdi%2BCVChjA7voJxvlDKq9suo%3D&amp;reserved=0
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>
> in the 'old days' in the core we used to cut filter paper and then bore a
> hole with a cork-bore slightly smaller than the lens.  slide 2-3 (or as
> many as will fit) layers of the filter paper over the objective.  this will
> not stop a "flood" of oil (which sounds very possible for some) but it will
> hold much more than you would think.  and, the most important feature of
> this little trick is that the oil will show very quickly on the paper and
> hopefully will trigger a response.!
>
> some of the darker filter papers will show the oil better, but this will
> vary by manufacturer.
>
> no commercial interest here, just trying to protect all the optics in the
> system.
>
> michael
>
> c. michael stanley, phd
> senior applications scientist
> chroma technology corp.
> 10 imtec lane
> bellows falls,  vt    05101
> [hidden email]
> [hidden email]
>
>
> -----Original Message-----
> From: Confocal Microscopy List <[hidden email]> On
> Behalf Of Gary Laevsky
> Sent: Tuesday, October 27, 2020 10:19 AM
> To: [hidden email]
> Subject: Re: Protective window for a confocal microscope?
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
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> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=WiaypCuZUL5s03B%2Br1rcmMLyOazUGYLj%2BmW7hRFcyIU%3D&amp;reserved=0
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> and include the link in your posting.
> *****
>
> The hair ties work great with objectives that have enough of a neck for
> the tie to sit on, next to the barrel.
>
> Our 40X objective (most frequently used) has no/very small neck, so the
> hair ties don't work.
>
> I fairly regularly (bi-weekly) have to take the objective off, store it in
> it's case upside down, and put it in a 37C oven.  Ridiculous, in a bad way,
> how much oil comes out.  The 40X also doesn't have "lip" that acts as a bit
> of a reservoir (unlike the 60 and 100).
>
> On one of my heavily used systems, I have to go so far as to dismantle the
> top (stage, objective turret, and fluorescent turret) to get to the piece
> of glass that prevents leakage into the body (amazingly awesome feature of
> the stand).  Takes me 15 minutes.  Although I shouldn't have to do that ...
>
> On Tue, Oct 27, 2020 at 9:13 AM Cammer, Michael <
> [hidden email]> wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists
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> > CI6Mn0%3D%7C1000&amp;sdata=q2oFBgmsbEfra%2BIBWB6i8xwEJ%2B8SF%2F59WSo3Y
> > %2BKvFKg%3D&amp;reserved=0 Post images on
> >
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> and include the link in your posting.
> > *****
> >
> > Aquastop works well for big spills when installed correctly and has
> > proven effective on some of our Zeiss scopes.  But we did have a user
> > who spilled over or around it on our Zeiss 710, so it is not foolproof.
> >
> >
> > Hair elastics are great for the oil.
> >
> >
> > Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory
> >
> > NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York,
> > NY
> > 10016
> >
> > [hidden email]<mailto:[hidden email]>
> > https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fnyulm
> > c.org%2Fmicros&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b8
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> > AN7BHwtFBKtNbXavROHTyX2m0CdnPe3l7VnUPA%3D&amp;reserved=0
> >
> > Voice direct only, no text or messages:  1-914-309-3270 and
> > 1-646-501-0567
> >
> >
> >
> > ________________________________
> > From: Confocal Microscopy List <[hidden email]> on
> > behalf of Doug Richardson <[hidden email]>
> > Sent: Tuesday, October 27, 2020 7:41:45 AM
> > To: [hidden email]
> > Subject: Re: Protective window for a confocal microscope?
> >
> > [EXTERNAL]
> >
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> >
> > https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-
> > 2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeEl
> > Zfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=ke
> > g1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=k-eCFZaqV2J3Nbh8c16znCG7uP
> > Pgwni6bBeSRyqYRAE&e=
> > Post images on
> > https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=Dw
> > IFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC5
> > 0tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-d
> > FQ&s=o9KAzzxXBHkEPM_YiZYo9mb7m2x6GjWRoUHh-BzcNSg&e=
> > and include the link in your posting.
> > *****
> >
> > I would highly recommend the aquastop system from Zeiss. It is
> > overpriced for some pieces of plastic, but it forms a very tight seal
> > and pays for itself many times over. It is far superior to hair
> > elastics/glove fingers/condoms which we used in the past.
> >
> > Doug
> >
> > Get Outlook for Android
> >
> > ________________________________
> > From: Confocal Microscopy List <[hidden email]> on
> > behalf of Tomek Węgierski <[hidden email]>
> > Sent: Tuesday, October 27, 2020 7:12:10 AM
> > To: [hidden email]
> > <[hidden email]>
> > Subject: Protective window for a confocal microscope?
> >
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> >
> > https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-
> > 2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeEl
> > Zfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=ke
> > g1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=k-eCFZaqV2J3Nbh8c16znCG7uP
> > Pgwni6bBeSRyqYRAE&e=
> > Post images on
> > https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=Dw
> > IFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC5
> > 0tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-d
> > FQ&s=o9KAzzxXBHkEPM_YiZYo9mb7m2x6GjWRoUHh-BzcNSg&e=
> > and include the link in your posting.
> > *****
> >
> > Dear All,
> > I am considering a purchase of a protective window for our Zeiss
> > LSM800 and I would like to hear your advice whether this is a good
> > idea, and if yes, which one to buy.
> > The reason is our LSM800 is on an inverted stand and we already got a
> > leakage of oil once which went down onto Optovar. And this was despite
> > my regular checks of objectives (both sides) for leaking oil and
> > extensive education of users on how important it is not to use too
> > much oil and to clean lenses, especially on inverted stands.
> > Zeiss Observer has a dummy slider with an opening for 32 mm filters. I
> > hope, a protective window there should stop the oil, if the leakage
> > happens again.
> > 1) do you think placing a protective window in a confocal microscope
> > is a good idea or not really, because the imaging will be affected?
> > This would be in infinity space, so I guess it should not be
> > detrimental....
> > 2) which characteristics of protective window I should take into
> > consideration (substrate, thickness, coating, flatness, surface
> > quality, parallelism)? Currently I am inclined into protective glasses
> > by Edmund Optics such as 1-mm thick MgF2-coated Sapphire window, which
> > has high transmission, or 2-mm thick MgF2-coated B270 window, which
> > has a bit worse transmission but better surface parameters (and is
> > much cheaper).
> > best regards,
> > Tomasz
> >
> >
> > Tomasz Wegierski, PhD
> > International Institute of Molecular and Cell Biology Trojdena 4,
> > 02-109 Warsaw, POLAND
> > tel: [phone]
> > fax: [phone]
> >
> > https://urldefense.proofpoint.com/v2/url?u=http-3A__www.iimcb.gov.pl_&
> > d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDi
> > PlC50tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNB
> > tS-dFQ&s=u7qHptX43byvVfMe-Zn04ev5KF4-ToW1ss01pq0eVtM&e=
> >
> > ------------------------------------------------------------
> > This email message, including any attachments, is for the sole use of
> > the intended recipient(s) and may contain information that is
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>
>
> --
> Best,
>
> Gary Laevsky, Ph.D.
> Director, Confocal Imaging Facility
> Nikon Center of Excellence
> Co-Founder, North Atlantic Microscopy Society (NAMS)
>
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--
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Zdenek Svindrych, Ph.D.
Research Scientist - Microscopy Imaging Specialist
Department of Biochemistry and Cell Biology
Geisel School of Medicine at Dartmouth

[Gary Laevsky]

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Hi All,

Staying on the question, in regards to putting a piece of glass in that
slider (which would make our lives easier), should it be placed on an
angle, to minimize reflections?  But, will that increase aberration?

Good question Tomasz.

On Tue, Oct 27, 2020 at 7:46 PM Zdenek Svindrych <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi Tomasz,
>
> back to your question. The filter holder is there to hold a filter. Any
> decent glass or fused silica AR window should do the trick.
> As far as I remember, the hole is quite big, around 30 mm diameter (bigger
> than the standard 25 mm filters), which may limit your options.
>
> Most filters and windows have specified surface flatness (reflected wave
> error), but it's actually transmitted wave error (TWE) that matters here.
> Usually, windows with single digit flatness (in lambda or fringe units)
> will have TWE << 1/5 lambda, and should be OK.
>
> Thicker substrate (2 - 3 mm) usually means precision optics, and is a safe
> bet. Hard coating is a must, if you want to wipe oil off of it regularly.
> All reputable brands use hard coatings these days, 1% loss (per pass) is
> OK.
>
> Sapphire is hard to work with and comes with a price premium. Fused silica,
> e.g.
>
> https://www.edmundoptics.com/p/30mm-dia-vis-nir-coated-1lambda-fused-silica-window/10158/
> (no commercial interest) looks like a good option, but BK-7 will work well,
> too.
>
> If you happen to have a piece of AR-coated glass at hand, just put it there
> and look at some beads. If you're not able to tell the difference, you're
> good to go. You can even try a glass coverslip, the 4% loss (the excitation
> loss is not an issue, you can always crank up the laser) will be hardly
> noticeable. Coverslips are not polished, so the TWE is generally bad, but
> with some luck you can find a good one in the stack...
>
> Best, zdenek
>
>
> On Tue, Oct 27, 2020 at 1:18 PM Tomek Węgierski <[hidden email]>
> wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > Dear All,
> >
> > Thank you very much for so many responses, although the discussion went a
> > bit into a direction that I did not anticipate ??
> > But yes, it is all about preventing oil from getting into the microscope.
> > Our 40x/1.3 Fluar lens takes a lot of oil and I frequently have to clean
> > its bottom side from oil. In contrast, I never saw any oil on the back of
> > 63x/1.4 even though it is used as frequently as the 40x.
> >
> > I have to admit I have been skeptical about hair elastics, because when
> > they saturate with oil they will just let it go. IMHO, it is replacing
> "oil
> > cleaning" approach with "oil buffering" approach. The "cleaning" approach
> > has worked for us for many years. It failed when an unexperienced user
> (who
> > apparently did not take seriously what I was talking during the training)
> > made two mistakes: did not clean the lens and let it stay in the optic
> path
> > after the work instead of changing for 10x. In addition, it was a holiday
> > time, the system was not used for the next few days, giving the oil
> plenty
> > of time. So, it was a coincidence of several factors, but it can happen
> > again.
> >
> > I think, hearing so many positive opinions about hair elastics, I will
> > give them the 2nd chance in our facility. Aqua-Stop in my opinion is
> > overpriced, and we do not currently do any experiments under perfusion to
> > justify such purchase (this may change in the future). However, I would
> not
> > like to give up on the protective window strategy. I think placing a
> glass
> > element in the infinity space is not unusual. It would function as the
> last
> > stronghold against the oil. And it would be considerably cheaper than
> > Aqua-Stop. I would of course test PSF. If PSF is not affected, I guess
> the
> > protective window is OK with imaging. So, if you still have some advice
> > which one would be most suitable, I would appreciate it.
> >
> > Best,
> > Tomasz
> >
> > Tomasz Wegierski, PhD
> > International Institute of Molecular and Cell Biology
> > Trojdena 4, 02-109 Warsaw, POLAND
> > tel: [phone]
> > fax: [phone]
> > http://www.iimcb.gov.pl/
> > ________________________________
> > From: Confocal Microscopy List <[hidden email]> on
> > behalf of Frohlich, Victoria <[hidden email]>
> > Sent: Tuesday, October 27, 2020 4:38 PM
> > To: [hidden email] <[hidden email]>
> > Subject: Re: Protective window for a confocal microscope?
> >
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > Just my 2 cents.
> > Scrunchies work well but are not fool-proof.  Cotton not polyester
> > material are more absorbent.
> > In addition to scrunchies I suggest wrapping the lens barrel near the
> nose
> > of the objective with plumbers tape to prevent oil from seeping into the
> > barrel.  It can be done so that the lens retraction still functions.
> Then
> > once the lens is on the nosepiece of the scope the plumbers tape can be
> > wrapped there as well.
> >
> > I've also used thin rubber sheets or parafilm sheets, as well as filter
> > paper (thanks Mike S.!), to draw oil away from the lens and nosepiece.
> >
> > Best method, however, is to teach users proper oil application and ensure
> > they clean up after themselves.
> >
> > Regards,
> > Vickie
> >
> > Victoria Centonze Frohlich, PhD
> > Director, Light Microscopy Shared Resource
> > Cell & Tissue Imaging Center
> > St. Jude Children’s Research Hospital
> > 262 Danny Thomas Place
> > MS 312
> > Memphis, TN 38105
> >
> > Office: D1052D
> > Phone and Digital Fax: [phone]
> > Cell: 1-901-907-2596
> > Paper Fax: [phone]
> > Email: [hidden email]
> >
> > CTIC-LM Location: D1055 and D1056
> > Facility Phone: [phone]
> >
> > http://home.stjude.org/cell-tissue-imaging/Pages/default.aspx
> >
> > Facility Acknowledgement for publications:
> > “Images were acquired at the Cell & Tissue Imaging Center which is
> > supported by SJCRH and NCI P30 CA021765.”
> >
> > -----Original Message-----
> > From: Confocal Microscopy List <[hidden email]> On
> > Behalf Of Michael Stanley
> > Sent: Tuesday, October 27, 2020 8:19 AM
> > To: [hidden email]
> > Subject: Re: Protective window for a confocal microscope?
> >
> > Caution: External Sender
> >
> >
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> >
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=WiaypCuZUL5s03B%2Br1rcmMLyOazUGYLj%2BmW7hRFcyIU%3D&amp;reserved=0
> > Post images on
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=DKQAcpfaPdTDony5B7X%2Fdi%2BCVChjA7voJxvlDKq9suo%3D&amp;reserved=0
> > and include the link in your posting.
> > *****
> >
> > in the 'old days' in the core we used to cut filter paper and then bore a
> > hole with a cork-bore slightly smaller than the lens.  slide 2-3 (or as
> > many as will fit) layers of the filter paper over the objective.  this
> will
> > not stop a "flood" of oil (which sounds very possible for some) but it
> will
> > hold much more than you would think.  and, the most important feature of
> > this little trick is that the oil will show very quickly on the paper and
> > hopefully will trigger a response.!
> >
> > some of the darker filter papers will show the oil better, but this will
> > vary by manufacturer.
> >
> > no commercial interest here, just trying to protect all the optics in the
> > system.
> >
> > michael
> >
> > c. michael stanley, phd
> > senior applications scientist
> > chroma technology corp.
> > 10 imtec lane
> > bellows falls,  vt    05101
> > [hidden email]
> > [hidden email]
> >
> >
> > -----Original Message-----
> > From: Confocal Microscopy List <[hidden email]> On
> > Behalf Of Gary Laevsky
> > Sent: Tuesday, October 27, 2020 10:19 AM
> > To: [hidden email]
> > Subject: Re: Protective window for a confocal microscope?
> >
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> >
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=WiaypCuZUL5s03B%2Br1rcmMLyOazUGYLj%2BmW7hRFcyIU%3D&amp;reserved=0
> > Post images on
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=yhWCT39l%2FiZJ9i8H0f4EM7MIstxliSvBVsjVQVNN8cI%3D&amp;reserved=0
> > and include the link in your posting.
> > *****
> >
> > The hair ties work great with objectives that have enough of a neck for
> > the tie to sit on, next to the barrel.
> >
> > Our 40X objective (most frequently used) has no/very small neck, so the
> > hair ties don't work.
> >
> > I fairly regularly (bi-weekly) have to take the objective off, store it
> in
> > it's case upside down, and put it in a 37C oven.  Ridiculous, in a bad
> way,
> > how much oil comes out.  The 40X also doesn't have "lip" that acts as a
> bit
> > of a reservoir (unlike the 60 and 100).
> >
> > On one of my heavily used systems, I have to go so far as to dismantle
> the
> > top (stage, objective turret, and fluorescent turret) to get to the piece
> > of glass that prevents leakage into the body (amazingly awesome feature
> of
> > the stand).  Takes me 15 minutes.  Although I shouldn't have to do that
> ...
> >
> > On Tue, Oct 27, 2020 at 9:13 AM Cammer, Michael <
> > [hidden email]> wrote:
> >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > > https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists
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> > > CI6Mn0%3D%7C1000&amp;sdata=q2oFBgmsbEfra%2BIBWB6i8xwEJ%2B8SF%2F59WSo3Y
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> > >
> >
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> > and include the link in your posting.
> > > *****
> > >
> > > Aquastop works well for big spills when installed correctly and has
> > > proven effective on some of our Zeiss scopes.  But we did have a user
> > > who spilled over or around it on our Zeiss 710, so it is not foolproof.
> > >
> > >
> > > Hair elastics are great for the oil.
> > >
> > >
> > > Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory
> > >
> > > NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York,
> > > NY
> > > 10016
> > >
> > > [hidden email]<mailto:[hidden email]>
> > > https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fnyulm
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> > >
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> > >
> > >
> > >
> > > ________________________________
> > > From: Confocal Microscopy List <[hidden email]> on
> > > behalf of Doug Richardson <[hidden email]>
> > > Sent: Tuesday, October 27, 2020 7:41:45 AM
> > > To: [hidden email]
> > > Subject: Re: Protective window for a confocal microscope?
> > >
> > > [EXTERNAL]
> > >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > >
> > > https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-
> > > 2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeEl
> > > Zfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=ke
> > > g1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=k-eCFZaqV2J3Nbh8c16znCG7uP
> > > Pgwni6bBeSRyqYRAE&e=
> > > Post images on
> > > https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=Dw
> > > IFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC5
> > > 0tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-d
> > > FQ&s=o9KAzzxXBHkEPM_YiZYo9mb7m2x6GjWRoUHh-BzcNSg&e=
> > > and include the link in your posting.
> > > *****
> > >
> > > I would highly recommend the aquastop system from Zeiss. It is
> > > overpriced for some pieces of plastic, but it forms a very tight seal
> > > and pays for itself many times over. It is far superior to hair
> > > elastics/glove fingers/condoms which we used in the past.
> > >
> > > Doug
> > >
> > > Get Outlook for Android
> > >
> > > ________________________________
> > > From: Confocal Microscopy List <[hidden email]> on
> > > behalf of Tomek Węgierski <[hidden email]>
> > > Sent: Tuesday, October 27, 2020 7:12:10 AM
> > > To: [hidden email]
> > > <[hidden email]>
> > > Subject: Protective window for a confocal microscope?
> > >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > >
> > > https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-
> > > 2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeEl
> > > Zfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=ke
> > > g1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=k-eCFZaqV2J3Nbh8c16znCG7uP
> > > Pgwni6bBeSRyqYRAE&e=
> > > Post images on
> > > https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=Dw
> > > IFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC5
> > > 0tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-d
> > > FQ&s=o9KAzzxXBHkEPM_YiZYo9mb7m2x6GjWRoUHh-BzcNSg&e=
> > > and include the link in your posting.
> > > *****
> > >
> > > Dear All,
> > > I am considering a purchase of a protective window for our Zeiss
> > > LSM800 and I would like to hear your advice whether this is a good
> > > idea, and if yes, which one to buy.
> > > The reason is our LSM800 is on an inverted stand and we already got a
> > > leakage of oil once which went down onto Optovar. And this was despite
> > > my regular checks of objectives (both sides) for leaking oil and
> > > extensive education of users on how important it is not to use too
> > > much oil and to clean lenses, especially on inverted stands.
> > > Zeiss Observer has a dummy slider with an opening for 32 mm filters. I
> > > hope, a protective window there should stop the oil, if the leakage
> > > happens again.
> > > 1) do you think placing a protective window in a confocal microscope
> > > is a good idea or not really, because the imaging will be affected?
> > > This would be in infinity space, so I guess it should not be
> > > detrimental....
> > > 2) which characteristics of protective window I should take into
> > > consideration (substrate, thickness, coating, flatness, surface
> > > quality, parallelism)? Currently I am inclined into protective glasses
> > > by Edmund Optics such as 1-mm thick MgF2-coated Sapphire window, which
> > > has high transmission, or 2-mm thick MgF2-coated B270 window, which
> > > has a bit worse transmission but better surface parameters (and is
> > > much cheaper).
> > > best regards,
> > > Tomasz
> > >
> > >
> > > Tomasz Wegierski, PhD
> > > International Institute of Molecular and Cell Biology Trojdena 4,
> > > 02-109 Warsaw, POLAND
> > > tel: [phone]
> > > fax: [phone]
> > >
> > > https://urldefense.proofpoint.com/v2/url?u=http-3A__www.iimcb.gov.pl_&
> > > d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDi
> > > PlC50tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNB
> > > tS-dFQ&s=u7qHptX43byvVfMe-Zn04ev5KF4-ToW1ss01pq0eVtM&e=
> > >
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> > > =================================
> > >
> >
> >
> > --
> > Best,
> >
> > Gary Laevsky, Ph.D.
> > Director, Confocal Imaging Facility
> > Nikon Center of Excellence
> > Co-Founder, North Atlantic Microscopy Society (NAMS)
> >
> >
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>
> --
> --
> Zdenek Svindrych, Ph.D.
> Research Scientist - Microscopy Imaging Specialist
> Department of Biochemistry and Cell Biology
> Geisel School of Medicine at Dartmouth
>

--
Best,

Gary Laevsky, Ph.D.
Director, Confocal Imaging Facility
Nikon Center of Excellence
Co-Founder, North Atlantic Microscopy Society (NAMS)
https://namsmicroscopy.com/
Dept. of Molecular Biology
Washington Rd.
Princeton University
Princeton, New Jersey, 08544-1014
(O) 609 258 5432
(C) 508 507 1310

[Tobias Baskin]

*****
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*****

Colleagues,
                     Given that the protective window would be on a
slider -- how difficult would it be to move into and out of position?
Perhaps for crtical optical applications, it would not be too difficult
to slide out of the beam? Stray thought.

Tobias

On 10/27/20 8:10 PM, Gary Laevsky wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi All,
>
> Staying on the question, in regards to putting a piece of glass in that
> slider (which would make our lives easier), should it be placed on an
> angle, to minimize reflections?  But, will that increase aberration?
>
> Good question Tomasz.
>
> On Tue, Oct 27, 2020 at 7:46 PM Zdenek Svindrych <[hidden email]> wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> Post images on http://www.imgur.com and include the link in your posting.
>> *****
>>
>> Hi Tomasz,
>>
>> back to your question. The filter holder is there to hold a filter. Any
>> decent glass or fused silica AR window should do the trick.
>> As far as I remember, the hole is quite big, around 30 mm diameter (bigger
>> than the standard 25 mm filters), which may limit your options.
>>
>> Most filters and windows have specified surface flatness (reflected wave
>> error), but it's actually transmitted wave error (TWE) that matters here.
>> Usually, windows with single digit flatness (in lambda or fringe units)
>> will have TWE << 1/5 lambda, and should be OK.
>>
>> Thicker substrate (2 - 3 mm) usually means precision optics, and is a safe
>> bet. Hard coating is a must, if you want to wipe oil off of it regularly.
>> All reputable brands use hard coatings these days, 1% loss (per pass) is
>> OK.
>>
>> Sapphire is hard to work with and comes with a price premium. Fused silica,
>> e.g.
>>
>> https://www.edmundoptics.com/p/30mm-dia-vis-nir-coated-1lambda-fused-silica-window/10158/
>> (no commercial interest) looks like a good option, but BK-7 will work well,
>> too.
>>
>> If you happen to have a piece of AR-coated glass at hand, just put it there
>> and look at some beads. If you're not able to tell the difference, you're
>> good to go. You can even try a glass coverslip, the 4% loss (the excitation
>> loss is not an issue, you can always crank up the laser) will be hardly
>> noticeable. Coverslips are not polished, so the TWE is generally bad, but
>> with some luck you can find a good one in the stack...
>>
>> Best, zdenek
>>
>>
>> On Tue, Oct 27, 2020 at 1:18 PM Tomek Węgierski <[hidden email]>
>> wrote:
>>
>>> *****
>>> To join, leave or search the confocal microscopy listserv, go to:
>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>> Post images on http://www.imgur.com and include the link in your
>> posting.
>>> *****
>>>
>>> Dear All,
>>>
>>> Thank you very much for so many responses, although the discussion went a
>>> bit into a direction that I did not anticipate ??
>>> But yes, it is all about preventing oil from getting into the microscope.
>>> Our 40x/1.3 Fluar lens takes a lot of oil and I frequently have to clean
>>> its bottom side from oil. In contrast, I never saw any oil on the back of
>>> 63x/1.4 even though it is used as frequently as the 40x.
>>>
>>> I have to admit I have been skeptical about hair elastics, because when
>>> they saturate with oil they will just let it go. IMHO, it is replacing
>> "oil
>>> cleaning" approach with "oil buffering" approach. The "cleaning" approach
>>> has worked for us for many years. It failed when an unexperienced user
>> (who
>>> apparently did not take seriously what I was talking during the training)
>>> made two mistakes: did not clean the lens and let it stay in the optic
>> path
>>> after the work instead of changing for 10x. In addition, it was a holiday
>>> time, the system was not used for the next few days, giving the oil
>> plenty
>>> of time. So, it was a coincidence of several factors, but it can happen
>>> again.
>>>
>>> I think, hearing so many positive opinions about hair elastics, I will
>>> give them the 2nd chance in our facility. Aqua-Stop in my opinion is
>>> overpriced, and we do not currently do any experiments under perfusion to
>>> justify such purchase (this may change in the future). However, I would
>> not
>>> like to give up on the protective window strategy. I think placing a
>> glass
>>> element in the infinity space is not unusual. It would function as the
>> last
>>> stronghold against the oil. And it would be considerably cheaper than
>>> Aqua-Stop. I would of course test PSF. If PSF is not affected, I guess
>> the
>>> protective window is OK with imaging. So, if you still have some advice
>>> which one would be most suitable, I would appreciate it.
>>>
>>> Best,
>>> Tomasz
>>>
>>> Tomasz Wegierski, PhD
>>> International Institute of Molecular and Cell Biology
>>> Trojdena 4, 02-109 Warsaw, POLAND
>>> tel: [phone]
>>> fax: [phone]
>>> http://www.iimcb.gov.pl/
>>> ________________________________
>>> From: Confocal Microscopy List <[hidden email]> on
>>> behalf of Frohlich, Victoria <[hidden email]>
>>> Sent: Tuesday, October 27, 2020 4:38 PM
>>> To: [hidden email] <[hidden email]>
>>> Subject: Re: Protective window for a confocal microscope?
>>>
>>> *****
>>> To join, leave or search the confocal microscopy listserv, go to:
>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>> Post images on http://www.imgur.com and include the link in your
>> posting.
>>> *****
>>>
>>> Just my 2 cents.
>>> Scrunchies work well but are not fool-proof.  Cotton not polyester
>>> material are more absorbent.
>>> In addition to scrunchies I suggest wrapping the lens barrel near the
>> nose
>>> of the objective with plumbers tape to prevent oil from seeping into the
>>> barrel.  It can be done so that the lens retraction still functions.
>> Then
>>> once the lens is on the nosepiece of the scope the plumbers tape can be
>>> wrapped there as well.
>>>
>>> I've also used thin rubber sheets or parafilm sheets, as well as filter
>>> paper (thanks Mike S.!), to draw oil away from the lens and nosepiece.
>>>
>>> Best method, however, is to teach users proper oil application and ensure
>>> they clean up after themselves.
>>>
>>> Regards,
>>> Vickie
>>>
>>> Victoria Centonze Frohlich, PhD
>>> Director, Light Microscopy Shared Resource
>>> Cell & Tissue Imaging Center
>>> St. Jude Children’s Research Hospital
>>> 262 Danny Thomas Place
>>> MS 312
>>> Memphis, TN 38105
>>>
>>> Office: D1052D
>>> Phone and Digital Fax: [phone]
>>> Cell: 1-901-907-2596
>>> Paper Fax: [phone]
>>> Email: [hidden email]
>>>
>>> CTIC-LM Location: D1055 and D1056
>>> Facility Phone: [phone]
>>>
>>> http://home.stjude.org/cell-tissue-imaging/Pages/default.aspx
>>>
>>> Facility Acknowledgement for publications:
>>> “Images were acquired at the Cell & Tissue Imaging Center which is
>>> supported by SJCRH and NCI P30 CA021765.”
>>>
>>> -----Original Message-----
>>> From: Confocal Microscopy List <[hidden email]> On
>>> Behalf Of Michael Stanley
>>> Sent: Tuesday, October 27, 2020 8:19 AM
>>> To: [hidden email]
>>> Subject: Re: Protective window for a confocal microscope?
>>>
>>> Caution: External Sender
>>>
>>>
>>> *****
>>> To join, leave or search the confocal microscopy listserv, go to:
>>>
>>>
>> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=WiaypCuZUL5s03B%2Br1rcmMLyOazUGYLj%2BmW7hRFcyIU%3D&amp;reserved=0
>>> Post images on
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>> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=DKQAcpfaPdTDony5B7X%2Fdi%2BCVChjA7voJxvlDKq9suo%3D&amp;reserved=0
>>> and include the link in your posting.
>>> *****
>>>
>>> in the 'old days' in the core we used to cut filter paper and then bore a
>>> hole with a cork-bore slightly smaller than the lens.  slide 2-3 (or as
>>> many as will fit) layers of the filter paper over the objective.  this
>> will
>>> not stop a "flood" of oil (which sounds very possible for some) but it
>> will
>>> hold much more than you would think.  and, the most important feature of
>>> this little trick is that the oil will show very quickly on the paper and
>>> hopefully will trigger a response.!
>>>
>>> some of the darker filter papers will show the oil better, but this will
>>> vary by manufacturer.
>>>
>>> no commercial interest here, just trying to protect all the optics in the
>>> system.
>>>
>>> michael
>>>
>>> c. michael stanley, phd
>>> senior applications scientist
>>> chroma technology corp.
>>> 10 imtec lane
>>> bellows falls,  vt    05101
>>> [hidden email]
>>> [hidden email]
>>>
>>>
>>> -----Original Message-----
>>> From: Confocal Microscopy List <[hidden email]> On
>>> Behalf Of Gary Laevsky
>>> Sent: Tuesday, October 27, 2020 10:19 AM
>>> To: [hidden email]
>>> Subject: Re: Protective window for a confocal microscope?
>>>
>>> *****
>>> To join, leave or search the confocal microscopy listserv, go to:
>>>
>>>
>> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=WiaypCuZUL5s03B%2Br1rcmMLyOazUGYLj%2BmW7hRFcyIU%3D&amp;reserved=0
>>> Post images on
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>> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=yhWCT39l%2FiZJ9i8H0f4EM7MIstxliSvBVsjVQVNN8cI%3D&amp;reserved=0
>>> and include the link in your posting.
>>> *****
>>>
>>> The hair ties work great with objectives that have enough of a neck for
>>> the tie to sit on, next to the barrel.
>>>
>>> Our 40X objective (most frequently used) has no/very small neck, so the
>>> hair ties don't work.
>>>
>>> I fairly regularly (bi-weekly) have to take the objective off, store it
>> in
>>> it's case upside down, and put it in a 37C oven.  Ridiculous, in a bad
>> way,
>>> how much oil comes out.  The 40X also doesn't have "lip" that acts as a
>> bit
>>> of a reservoir (unlike the 60 and 100).
>>>
>>> On one of my heavily used systems, I have to go so far as to dismantle
>> the
>>> top (stage, objective turret, and fluorescent turret) to get to the piece
>>> of glass that prevents leakage into the body (amazingly awesome feature
>> of
>>> the stand).  Takes me 15 minutes.  Although I shouldn't have to do that
>> ...
>>> On Tue, Oct 27, 2020 at 9:13 AM Cammer, Michael <
>>> [hidden email]> wrote:
>>>
>>>> *****
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>>> and include the link in your posting.
>>>> *****
>>>>
>>>> Aquastop works well for big spills when installed correctly and has
>>>> proven effective on some of our Zeiss scopes.  But we did have a user
>>>> who spilled over or around it on our Zeiss 710, so it is not foolproof.
>>>>
>>>>
>>>> Hair elastics are great for the oil.
>>>>
>>>>
>>>> Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory
>>>>
>>>> NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York,
>>>> NY
>>>> 10016
>>>>
>>>> [hidden email]<mailto:[hidden email]>
>>>> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fnyulm
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>>>> JQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=Ud4O3pAe
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>>>>
>>>> Voice direct only, no text or messages:  1-914-309-3270 and
>>>> 1-646-501-0567
>>>>
>>>>
>>>>
>>>> ________________________________
>>>> From: Confocal Microscopy List <[hidden email]> on
>>>> behalf of Doug Richardson <[hidden email]>
>>>> Sent: Tuesday, October 27, 2020 7:41:45 AM
>>>> To: [hidden email]
>>>> Subject: Re: Protective window for a confocal microscope?
>>>>
>>>> [EXTERNAL]
>>>>
>>>> *****
>>>> To join, leave or search the confocal microscopy listserv, go to:
>>>>
>>>> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-
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>>>> Zfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=ke
>>>> g1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=k-eCFZaqV2J3Nbh8c16znCG7uP
>>>> Pgwni6bBeSRyqYRAE&e=
>>>> Post images on
>>>> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=Dw
>>>> IFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC5
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>>>> and include the link in your posting.
>>>> *****
>>>>
>>>> I would highly recommend the aquastop system from Zeiss. It is
>>>> overpriced for some pieces of plastic, but it forms a very tight seal
>>>> and pays for itself many times over. It is far superior to hair
>>>> elastics/glove fingers/condoms which we used in the past.
>>>>
>>>> Doug
>>>>
>>>> Get Outlook for Android
>>>>
>>>> ________________________________
>>>> From: Confocal Microscopy List <[hidden email]> on
>>>> behalf of Tomek Węgierski <[hidden email]>
>>>> Sent: Tuesday, October 27, 2020 7:12:10 AM
>>>> To: [hidden email]
>>>> <[hidden email]>
>>>> Subject: Protective window for a confocal microscope?
>>>>
>>>> *****
>>>> To join, leave or search the confocal microscopy listserv, go to:
>>>>
>>>> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-
>>>> 2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeEl
>>>> Zfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=ke
>>>> g1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=k-eCFZaqV2J3Nbh8c16znCG7uP
>>>> Pgwni6bBeSRyqYRAE&e=
>>>> Post images on
>>>> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=Dw
>>>> IFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC5
>>>> 0tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-d
>>>> FQ&s=o9KAzzxXBHkEPM_YiZYo9mb7m2x6GjWRoUHh-BzcNSg&e=
>>>> and include the link in your posting.
>>>> *****
>>>>
>>>> Dear All,
>>>> I am considering a purchase of a protective window for our Zeiss
>>>> LSM800 and I would like to hear your advice whether this is a good
>>>> idea, and if yes, which one to buy.
>>>> The reason is our LSM800 is on an inverted stand and we already got a
>>>> leakage of oil once which went down onto Optovar. And this was despite
>>>> my regular checks of objectives (both sides) for leaking oil and
>>>> extensive education of users on how important it is not to use too
>>>> much oil and to clean lenses, especially on inverted stands.
>>>> Zeiss Observer has a dummy slider with an opening for 32 mm filters. I
>>>> hope, a protective window there should stop the oil, if the leakage
>>>> happens again.
>>>> 1) do you think placing a protective window in a confocal microscope
>>>> is a good idea or not really, because the imaging will be affected?
>>>> This would be in infinity space, so I guess it should not be
>>>> detrimental....
>>>> 2) which characteristics of protective window I should take into
>>>> consideration (substrate, thickness, coating, flatness, surface
>>>> quality, parallelism)? Currently I am inclined into protective glasses
>>>> by Edmund Optics such as 1-mm thick MgF2-coated Sapphire window, which
>>>> has high transmission, or 2-mm thick MgF2-coated B270 window, which
>>>> has a bit worse transmission but better surface parameters (and is
>>>> much cheaper).
>>>> best regards,
>>>> Tomasz
>>>>
>>>>
>>>> Tomasz Wegierski, PhD
>>>> International Institute of Molecular and Cell Biology Trojdena 4,
>>>> 02-109 Warsaw, POLAND
>>>> tel: [phone]
>>>> fax: [phone]
>>>>
>>>> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.iimcb.gov.pl_&
>>>> d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDi
>>>> PlC50tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNB
>>>> tS-dFQ&s=u7qHptX43byvVfMe-Zn04ev5KF4-ToW1ss01pq0eVtM&e=
>>>>
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>>>>
>>> --
>>> Best,
>>>
>>> Gary Laevsky, Ph.D.
>>> Director, Confocal Imaging Facility
>>> Nikon Center of Excellence
>>> Co-Founder, North Atlantic Microscopy Society (NAMS)
>>>
>>>
>> https://nam11.safelinks.protection.outlook.com/?url=https%3A%2F%2Fnamsmicroscopy.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=7foXnZ0OuaU32Fj2dlCniXz%2Bn8iWTBvMyOkGj0rTX10%3D&amp;reserved=0
>>> Dept. of Molecular Biology Washington Rd.
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>>> (O) 609 258 5432
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>>>
>>> ________________________________
>>>
>>> Email Disclaimer: www.stjude.org/emaildisclaimer<
>>> http://www.stjude.org/emaildisclaimer>
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>>> http://www.stjude.org/consultationdisclaimer>
>>>
>> --
>> --
>> Zdenek Svindrych, Ph.D.
>> Research Scientist - Microscopy Imaging Specialist
>> Department of Biochemistry and Cell Biology
>> Geisel School of Medicine at Dartmouth
>>

.     __    ___   ^    ___   ___   Tobias I. Baskin
      /  \  /     / \  /      \      Professor   (he)
     /   / /     /   \ \       \      Biology Department
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[Zdenek Svindrych-2]

*****
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Post images on http://www.imgur.com and include the link in your posting.
*****

Hi,

most microscopes contain some sort of angled beam splitter or dichroic
filter in the infinity space. And, indeed, most flat surfaces (windows,
filters) are angled.

zdenek


On Tue, Oct 27, 2020 at 8:11 PM Gary Laevsky <[hidden email]>
wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi All,
>
> Staying on the question, in regards to putting a piece of glass in that
> slider (which would make our lives easier), should it be placed on an
> angle, to minimize reflections?  But, will that increase aberration?
>
> Good question Tomasz.
>
> On Tue, Oct 27, 2020 at 7:46 PM Zdenek Svindrych <[hidden email]>
> wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > Hi Tomasz,
> >
> > back to your question. The filter holder is there to hold a filter. Any
> > decent glass or fused silica AR window should do the trick.
> > As far as I remember, the hole is quite big, around 30 mm diameter
> (bigger
> > than the standard 25 mm filters), which may limit your options.
> >
> > Most filters and windows have specified surface flatness (reflected wave
> > error), but it's actually transmitted wave error (TWE) that matters here.
> > Usually, windows with single digit flatness (in lambda or fringe units)
> > will have TWE << 1/5 lambda, and should be OK.
> >
> > Thicker substrate (2 - 3 mm) usually means precision optics, and is a
> safe
> > bet. Hard coating is a must, if you want to wipe oil off of it regularly.
> > All reputable brands use hard coatings these days, 1% loss (per pass) is
> > OK.
> >
> > Sapphire is hard to work with and comes with a price premium. Fused
> silica,
> > e.g.
> >
> >
> https://www.edmundoptics.com/p/30mm-dia-vis-nir-coated-1lambda-fused-silica-window/10158/
> > (no commercial interest) looks like a good option, but BK-7 will work
> well,
> > too.
> >
> > If you happen to have a piece of AR-coated glass at hand, just put it
> there
> > and look at some beads. If you're not able to tell the difference, you're
> > good to go. You can even try a glass coverslip, the 4% loss (the
> excitation
> > loss is not an issue, you can always crank up the laser) will be hardly
> > noticeable. Coverslips are not polished, so the TWE is generally bad, but
> > with some luck you can find a good one in the stack...
> >
> > Best, zdenek
> >
> >
> > On Tue, Oct 27, 2020 at 1:18 PM Tomek Węgierski <[hidden email]>
> > wrote:
> >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > > Post images on http://www.imgur.com and include the link in your
> > posting.
> > > *****
> > >
> > > Dear All,
> > >
> > > Thank you very much for so many responses, although the discussion
> went a
> > > bit into a direction that I did not anticipate ??
> > > But yes, it is all about preventing oil from getting into the
> microscope.
> > > Our 40x/1.3 Fluar lens takes a lot of oil and I frequently have to
> clean
> > > its bottom side from oil. In contrast, I never saw any oil on the back
> of
> > > 63x/1.4 even though it is used as frequently as the 40x.
> > >
> > > I have to admit I have been skeptical about hair elastics, because when
> > > they saturate with oil they will just let it go. IMHO, it is replacing
> > "oil
> > > cleaning" approach with "oil buffering" approach. The "cleaning"
> approach
> > > has worked for us for many years. It failed when an unexperienced user
> > (who
> > > apparently did not take seriously what I was talking during the
> training)
> > > made two mistakes: did not clean the lens and let it stay in the optic
> > path
> > > after the work instead of changing for 10x. In addition, it was a
> holiday
> > > time, the system was not used for the next few days, giving the oil
> > plenty
> > > of time. So, it was a coincidence of several factors, but it can happen
> > > again.
> > >
> > > I think, hearing so many positive opinions about hair elastics, I will
> > > give them the 2nd chance in our facility. Aqua-Stop in my opinion is
> > > overpriced, and we do not currently do any experiments under perfusion
> to
> > > justify such purchase (this may change in the future). However, I would
> > not
> > > like to give up on the protective window strategy. I think placing a
> > glass
> > > element in the infinity space is not unusual. It would function as the
> > last
> > > stronghold against the oil. And it would be considerably cheaper than
> > > Aqua-Stop. I would of course test PSF. If PSF is not affected, I guess
> > the
> > > protective window is OK with imaging. So, if you still have some advice
> > > which one would be most suitable, I would appreciate it.
> > >
> > > Best,
> > > Tomasz
> > >
> > > Tomasz Wegierski, PhD
> > > International Institute of Molecular and Cell Biology
> > > Trojdena 4, 02-109 Warsaw, POLAND
> > > tel: [phone]
> > > fax: [phone]
> > > http://www.iimcb.gov.pl/
> > > ________________________________
> > > From: Confocal Microscopy List <[hidden email]> on
> > > behalf of Frohlich, Victoria <[hidden email]>
> > > Sent: Tuesday, October 27, 2020 4:38 PM
> > > To: [hidden email] <[hidden email]
> >
> > > Subject: Re: Protective window for a confocal microscope?
> > >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > > Post images on http://www.imgur.com and include the link in your
> > posting.
> > > *****
> > >
> > > Just my 2 cents.
> > > Scrunchies work well but are not fool-proof.  Cotton not polyester
> > > material are more absorbent.
> > > In addition to scrunchies I suggest wrapping the lens barrel near the
> > nose
> > > of the objective with plumbers tape to prevent oil from seeping into
> the
> > > barrel.  It can be done so that the lens retraction still functions.
> > Then
> > > once the lens is on the nosepiece of the scope the plumbers tape can be
> > > wrapped there as well.
> > >
> > > I've also used thin rubber sheets or parafilm sheets, as well as filter
> > > paper (thanks Mike S.!), to draw oil away from the lens and nosepiece.
> > >
> > > Best method, however, is to teach users proper oil application and
> ensure
> > > they clean up after themselves.
> > >
> > > Regards,
> > > Vickie
> > >
> > > Victoria Centonze Frohlich, PhD
> > > Director, Light Microscopy Shared Resource
> > > Cell & Tissue Imaging Center
> > > St. Jude Children’s Research Hospital
> > > 262 Danny Thomas Place
> > > MS 312
> > > Memphis, TN 38105
> > >
> > > Office: D1052D
> > > Phone and Digital Fax: [phone]
> > > Cell: 1-901-907-2596
> > > Paper Fax: [phone]
> > > Email: [hidden email]
> > >
> > > CTIC-LM Location: D1055 and D1056
> > > Facility Phone: [phone]
> > >
> > > http://home.stjude.org/cell-tissue-imaging/Pages/default.aspx
> > >
> > > Facility Acknowledgement for publications:
> > > “Images were acquired at the Cell & Tissue Imaging Center which is
> > > supported by SJCRH and NCI P30 CA021765.”
> > >
> > > -----Original Message-----
> > > From: Confocal Microscopy List <[hidden email]> On
> > > Behalf Of Michael Stanley
> > > Sent: Tuesday, October 27, 2020 8:19 AM
> > > To: [hidden email]
> > > Subject: Re: Protective window for a confocal microscope?
> > >
> > > Caution: External Sender
> > >
> > >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > >
> > >
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=WiaypCuZUL5s03B%2Br1rcmMLyOazUGYLj%2BmW7hRFcyIU%3D&amp;reserved=0
> > > Post images on
> > >
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=DKQAcpfaPdTDony5B7X%2Fdi%2BCVChjA7voJxvlDKq9suo%3D&amp;reserved=0
> > > and include the link in your posting.
> > > *****
> > >
> > > in the 'old days' in the core we used to cut filter paper and then
> bore a
> > > hole with a cork-bore slightly smaller than the lens.  slide 2-3 (or as
> > > many as will fit) layers of the filter paper over the objective.  this
> > will
> > > not stop a "flood" of oil (which sounds very possible for some) but it
> > will
> > > hold much more than you would think.  and, the most important feature
> of
> > > this little trick is that the oil will show very quickly on the paper
> and
> > > hopefully will trigger a response.!
> > >
> > > some of the darker filter papers will show the oil better, but this
> will
> > > vary by manufacturer.
> > >
> > > no commercial interest here, just trying to protect all the optics in
> the
> > > system.
> > >
> > > michael
> > >
> > > c. michael stanley, phd
> > > senior applications scientist
> > > chroma technology corp.
> > > 10 imtec lane
> > > bellows falls,  vt    05101
> > > [hidden email]
> > > [hidden email]
> > >
> > >
> > > -----Original Message-----
> > > From: Confocal Microscopy List <[hidden email]> On
> > > Behalf Of Gary Laevsky
> > > Sent: Tuesday, October 27, 2020 10:19 AM
> > > To: [hidden email]
> > > Subject: Re: Protective window for a confocal microscope?
> > >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > >
> > >
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=WiaypCuZUL5s03B%2Br1rcmMLyOazUGYLj%2BmW7hRFcyIU%3D&amp;reserved=0
> > > Post images on
> > >
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=yhWCT39l%2FiZJ9i8H0f4EM7MIstxliSvBVsjVQVNN8cI%3D&amp;reserved=0
> > > and include the link in your posting.
> > > *****
> > >
> > > The hair ties work great with objectives that have enough of a neck for
> > > the tie to sit on, next to the barrel.
> > >
> > > Our 40X objective (most frequently used) has no/very small neck, so the
> > > hair ties don't work.
> > >
> > > I fairly regularly (bi-weekly) have to take the objective off, store it
> > in
> > > it's case upside down, and put it in a 37C oven.  Ridiculous, in a bad
> > way,
> > > how much oil comes out.  The 40X also doesn't have "lip" that acts as a
> > bit
> > > of a reservoir (unlike the 60 and 100).
> > >
> > > On one of my heavily used systems, I have to go so far as to dismantle
> > the
> > > top (stage, objective turret, and fluorescent turret) to get to the
> piece
> > > of glass that prevents leakage into the body (amazingly awesome feature
> > of
> > > the stand).  Takes me 15 minutes.  Although I shouldn't have to do that
> > ...
> > >
> > > On Tue, Oct 27, 2020 at 9:13 AM Cammer, Michael <
> > > [hidden email]> wrote:
> > >
> > > > *****
> > > > To join, leave or search the confocal microscopy listserv, go to:
> > > >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists
> > > > .umn.edu
> %2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=04%7C01%7Cv
> > > > ictoria.frohlich%40STJUDE.ORG
> %7C6b89a1a0f4cc42f5937e08d87a8d1047%7C223
> > > >
> 40fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573459000%7CUnknown%7
> > > >
> CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXV
> > > >
> CI6Mn0%3D%7C1000&amp;sdata=q2oFBgmsbEfra%2BIBWB6i8xwEJ%2B8SF%2F59WSo3Y
> > > > %2BKvFKg%3D&amp;reserved=0 Post images on
> > > >
> > >
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=yhWCT39l%2FiZJ9i8H0f4EM7MIstxliSvBVsjVQVNN8cI%3D&amp;reserved=0
> > > and include the link in your posting.
> > > > *****
> > > >
> > > > Aquastop works well for big spills when installed correctly and has
> > > > proven effective on some of our Zeiss scopes.  But we did have a user
> > > > who spilled over or around it on our Zeiss 710, so it is not
> foolproof.
> > > >
> > > >
> > > > Hair elastics are great for the oil.
> > > >
> > > >
> > > > Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory
> > > >
> > > > NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York,
> > > > NY
> > > > 10016
> > > >
> > > > [hidden email]<mailto:[hidden email]>
> > > >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fnyulm
> > > > c.org%2Fmicros&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG
> %7C6b8
> > > >
> 9a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7
> > > >
> C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLC
> > > >
> JQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=Ud4O3pAe
> > > > 6c4HlzFgcjkQBvmAAz45ILNKOj8mnQScAsg%3D&amp;reserved=0
> > > >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fmicro
> > > > scopynotes.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG
> %7C
> > > >
> 6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C
> > > >
> 0%7C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDA
> > > >
> iLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=e6srs
> > > > AN7BHwtFBKtNbXavROHTyX2m0CdnPe3l7VnUPA%3D&amp;reserved=0
> > > >
> > > > Voice direct only, no text or messages:  1-914-309-3270 and
> > > > 1-646-501-0567
> > > >
> > > >
> > > >
> > > > ________________________________
> > > > From: Confocal Microscopy List <[hidden email]> on
> > > > behalf of Doug Richardson <[hidden email]>
> > > > Sent: Tuesday, October 27, 2020 7:41:45 AM
> > > > To: [hidden email]
> > > > Subject: Re: Protective window for a confocal microscope?
> > > >
> > > > [EXTERNAL]
> > > >
> > > > *****
> > > > To join, leave or search the confocal microscopy listserv, go to:
> > > >
> > > >
> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-
> > > >
> 2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeEl
> > > >
> Zfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=ke
> > > >
> g1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=k-eCFZaqV2J3Nbh8c16znCG7uP
> > > > Pgwni6bBeSRyqYRAE&e=
> > > > Post images on
> > > >
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=Dw
> > > >
> IFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC5
> > > >
> 0tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-d
> > > > FQ&s=o9KAzzxXBHkEPM_YiZYo9mb7m2x6GjWRoUHh-BzcNSg&e=
> > > > and include the link in your posting.
> > > > *****
> > > >
> > > > I would highly recommend the aquastop system from Zeiss. It is
> > > > overpriced for some pieces of plastic, but it forms a very tight seal
> > > > and pays for itself many times over. It is far superior to hair
> > > > elastics/glove fingers/condoms which we used in the past.
> > > >
> > > > Doug
> > > >
> > > > Get Outlook for Android
> > > >
> > > > ________________________________
> > > > From: Confocal Microscopy List <[hidden email]> on
> > > > behalf of Tomek Węgierski <[hidden email]>
> > > > Sent: Tuesday, October 27, 2020 7:12:10 AM
> > > > To: [hidden email]
> > > > <[hidden email]>
> > > > Subject: Protective window for a confocal microscope?
> > > >
> > > > *****
> > > > To join, leave or search the confocal microscopy listserv, go to:
> > > >
> > > >
> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-
> > > >
> 2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeEl
> > > >
> Zfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=ke
> > > >
> g1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=k-eCFZaqV2J3Nbh8c16znCG7uP
> > > > Pgwni6bBeSRyqYRAE&e=
> > > > Post images on
> > > >
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=Dw
> > > >
> IFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC5
> > > >
> 0tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-d
> > > > FQ&s=o9KAzzxXBHkEPM_YiZYo9mb7m2x6GjWRoUHh-BzcNSg&e=
> > > > and include the link in your posting.
> > > > *****
> > > >
> > > > Dear All,
> > > > I am considering a purchase of a protective window for our Zeiss
> > > > LSM800 and I would like to hear your advice whether this is a good
> > > > idea, and if yes, which one to buy.
> > > > The reason is our LSM800 is on an inverted stand and we already got a
> > > > leakage of oil once which went down onto Optovar. And this was
> despite
> > > > my regular checks of objectives (both sides) for leaking oil and
> > > > extensive education of users on how important it is not to use too
> > > > much oil and to clean lenses, especially on inverted stands.
> > > > Zeiss Observer has a dummy slider with an opening for 32 mm filters.
> I
> > > > hope, a protective window there should stop the oil, if the leakage
> > > > happens again.
> > > > 1) do you think placing a protective window in a confocal microscope
> > > > is a good idea or not really, because the imaging will be affected?
> > > > This would be in infinity space, so I guess it should not be
> > > > detrimental....
> > > > 2) which characteristics of protective window I should take into
> > > > consideration (substrate, thickness, coating, flatness, surface
> > > > quality, parallelism)? Currently I am inclined into protective
> glasses
> > > > by Edmund Optics such as 1-mm thick MgF2-coated Sapphire window,
> which
> > > > has high transmission, or 2-mm thick MgF2-coated B270 window, which
> > > > has a bit worse transmission but better surface parameters (and is
> > > > much cheaper).
> > > > best regards,
> > > > Tomasz
> > > >
> > > >
> > > > Tomasz Wegierski, PhD
> > > > International Institute of Molecular and Cell Biology Trojdena 4,
> > > > 02-109 Warsaw, POLAND
> > > > tel: [phone]
> > > > fax: [phone]
> > > >
> > > >
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.iimcb.gov.pl_&
> > > >
> d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDi
> > > >
> PlC50tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNB
> > > > tS-dFQ&s=u7qHptX43byvVfMe-Zn04ev5KF4-ToW1ss01pq0eVtM&e=
> > > >
> > > > ------------------------------------------------------------
> > > > This email message, including any attachments, is for the sole use of
> > > > the intended recipient(s) and may contain information that is
> > > > proprietary, confidential, and exempt from disclosure under
> applicable
> > > > law. Any unauthorized review, use, disclosure, or distribution is
> > > > prohibited. If you have received this email in error please notify
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> > > > damage caused by any virus transmitted by this email.
> > > > =================================
> > > >
> > >
> > >
> > > --
> > > Best,
> > >
> > > Gary Laevsky, Ph.D.
> > > Director, Confocal Imaging Facility
> > > Nikon Center of Excellence
> > > Co-Founder, North Atlantic Microscopy Society (NAMS)
> > >
> > >
> >
> https://nam11.safelinks.protection.outlook.com/?url=https%3A%2F%2Fnamsmicroscopy.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=7foXnZ0OuaU32Fj2dlCniXz%2Bn8iWTBvMyOkGj0rTX10%3D&amp;reserved=0
> > > Dept. of Molecular Biology Washington Rd.
> > > Princeton University
> > > Princeton, New Jersey, 08544-1014
> > > (O) 609 258 5432
> > > (C) 508 507 1310
> > >
> > > ________________________________
> > >
> > > Email Disclaimer: www.stjude.org/emaildisclaimer<
> > > http://www.stjude.org/emaildisclaimer>
> > > Consultation Disclaimer: www.stjude.org/consultationdisclaimer<
> > > http://www.stjude.org/consultationdisclaimer>
> > >
> >
> >
> > --
> > --
> > Zdenek Svindrych, Ph.D.
> > Research Scientist - Microscopy Imaging Specialist
> > Department of Biochemistry and Cell Biology
> > Geisel School of Medicine at Dartmouth
> >
>
>
> --
> Best,
>
> Gary Laevsky, Ph.D.
> Director, Confocal Imaging Facility
> Nikon Center of Excellence
> Co-Founder, North Atlantic Microscopy Society (NAMS)
> https://namsmicroscopy.com/
> Dept. of Molecular Biology
> Washington Rd.
> Princeton University
> Princeton, New Jersey, 08544-1014
> (O) 609 258 5432
> (C) 508 507 1310
>

--
--
Zdenek Svindrych, Ph.D.
Research Scientist - Microscopy Imaging Specialist
Department of Biochemistry and Cell Biology
Geisel School of Medicine at Dartmouth

[Tomek Węgierski]

*****
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Post images on http://www.imgur.com and include the link in your posting.
*****

Dear All,
Following advice of Zdenek (thank you!) on the importance of TWE I decided to order this:
https://www.edmundoptics.de/p/30mm-dia-4mm-thick-vis-0deg-coated-lambda10-fused-silica-window/7997/
Its transmitted wavefront is lambda/10, and it shows very good parallelism, surface finish and transmission for VIS light, which is sufficient for us.
I guess the opening for filters in the filter slider is angled, but I will check this. And the slider can be removed anytime from the optic path in no time, so it will be very easy to check the imaging with and without the protective window.
When I test the PSFs I will let you know about the results.
Best,
Tomasz
PS. In my previous mail I meant 40x/1.3 EC Plan-Neofluar lens rather than "Fluar", which is a different lens (and less suitable for confocal). So, I meant one thing but somehow wrote the other. Sorry for that.

Tomasz Wegierski, PhD
International Institute of Molecular and Cell Biology
Trojdena 4, 02-109 Warsaw, POLAND
tel: [phone]
fax: [phone]
http://www.iimcb.gov.pl/
________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Zdenek Svindrych <[hidden email]>
Sent: Wednesday, October 28, 2020 12:50 PM
To: [hidden email] <[hidden email]>
Subject: Re: Protective window for a confocal microscope?

*****
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Post images on http://www.imgur.com and include the link in your posting.
*****

Hi,

most microscopes contain some sort of angled beam splitter or dichroic
filter in the infinity space. And, indeed, most flat surfaces (windows,
filters) are angled.

zdenek


On Tue, Oct 27, 2020 at 8:11 PM Gary Laevsky <[hidden email]>
wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi All,
>
> Staying on the question, in regards to putting a piece of glass in that
> slider (which would make our lives easier), should it be placed on an
> angle, to minimize reflections?  But, will that increase aberration?
>
> Good question Tomasz.
>
> On Tue, Oct 27, 2020 at 7:46 PM Zdenek Svindrych <[hidden email]>
> wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > Hi Tomasz,
> >
> > back to your question. The filter holder is there to hold a filter. Any
> > decent glass or fused silica AR window should do the trick.
> > As far as I remember, the hole is quite big, around 30 mm diameter
> (bigger
> > than the standard 25 mm filters), which may limit your options.
> >
> > Most filters and windows have specified surface flatness (reflected wave
> > error), but it's actually transmitted wave error (TWE) that matters here.
> > Usually, windows with single digit flatness (in lambda or fringe units)
> > will have TWE << 1/5 lambda, and should be OK.
> >
> > Thicker substrate (2 - 3 mm) usually means precision optics, and is a
> safe
> > bet. Hard coating is a must, if you want to wipe oil off of it regularly.
> > All reputable brands use hard coatings these days, 1% loss (per pass) is
> > OK.
> >
> > Sapphire is hard to work with and comes with a price premium. Fused
> silica,
> > e.g.
> >
> >
> https://www.edmundoptics.com/p/30mm-dia-vis-nir-coated-1lambda-fused-silica-window/10158/
> > (no commercial interest) looks like a good option, but BK-7 will work
> well,
> > too.
> >
> > If you happen to have a piece of AR-coated glass at hand, just put it
> there
> > and look at some beads. If you're not able to tell the difference, you're
> > good to go. You can even try a glass coverslip, the 4% loss (the
> excitation
> > loss is not an issue, you can always crank up the laser) will be hardly
> > noticeable. Coverslips are not polished, so the TWE is generally bad, but
> > with some luck you can find a good one in the stack...
> >
> > Best, zdenek
> >
> >
> > On Tue, Oct 27, 2020 at 1:18 PM Tomek Węgierski <[hidden email]>
> > wrote:
> >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > > Post images on http://www.imgur.com and include the link in your
> > posting.
> > > *****
> > >
> > > Dear All,
> > >
> > > Thank you very much for so many responses, although the discussion
> went a
> > > bit into a direction that I did not anticipate ??
> > > But yes, it is all about preventing oil from getting into the
> microscope.
> > > Our 40x/1.3 Fluar lens takes a lot of oil and I frequently have to
> clean
> > > its bottom side from oil. In contrast, I never saw any oil on the back
> of
> > > 63x/1.4 even though it is used as frequently as the 40x.
> > >
> > > I have to admit I have been skeptical about hair elastics, because when
> > > they saturate with oil they will just let it go. IMHO, it is replacing
> > "oil
> > > cleaning" approach with "oil buffering" approach. The "cleaning"
> approach
> > > has worked for us for many years. It failed when an unexperienced user
> > (who
> > > apparently did not take seriously what I was talking during the
> training)
> > > made two mistakes: did not clean the lens and let it stay in the optic
> > path
> > > after the work instead of changing for 10x. In addition, it was a
> holiday
> > > time, the system was not used for the next few days, giving the oil
> > plenty
> > > of time. So, it was a coincidence of several factors, but it can happen
> > > again.
> > >
> > > I think, hearing so many positive opinions about hair elastics, I will
> > > give them the 2nd chance in our facility. Aqua-Stop in my opinion is
> > > overpriced, and we do not currently do any experiments under perfusion
> to
> > > justify such purchase (this may change in the future). However, I would
> > not
> > > like to give up on the protective window strategy. I think placing a
> > glass
> > > element in the infinity space is not unusual. It would function as the
> > last
> > > stronghold against the oil. And it would be considerably cheaper than
> > > Aqua-Stop. I would of course test PSF. If PSF is not affected, I guess
> > the
> > > protective window is OK with imaging. So, if you still have some advice
> > > which one would be most suitable, I would appreciate it.
> > >
> > > Best,
> > > Tomasz
> > >
> > > Tomasz Wegierski, PhD
> > > International Institute of Molecular and Cell Biology
> > > Trojdena 4, 02-109 Warsaw, POLAND
> > > tel: [phone]
> > > fax: [phone]
> > > http://www.iimcb.gov.pl/
> > > ________________________________
> > > From: Confocal Microscopy List <[hidden email]> on
> > > behalf of Frohlich, Victoria <[hidden email]>
> > > Sent: Tuesday, October 27, 2020 4:38 PM
> > > To: [hidden email] <[hidden email]
> >
> > > Subject: Re: Protective window for a confocal microscope?
> > >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > > Post images on http://www.imgur.com and include the link in your
> > posting.
> > > *****
> > >
> > > Just my 2 cents.
> > > Scrunchies work well but are not fool-proof.  Cotton not polyester
> > > material are more absorbent.
> > > In addition to scrunchies I suggest wrapping the lens barrel near the
> > nose
> > > of the objective with plumbers tape to prevent oil from seeping into
> the
> > > barrel.  It can be done so that the lens retraction still functions.
> > Then
> > > once the lens is on the nosepiece of the scope the plumbers tape can be
> > > wrapped there as well.
> > >
> > > I've also used thin rubber sheets or parafilm sheets, as well as filter
> > > paper (thanks Mike S.!), to draw oil away from the lens and nosepiece.
> > >
> > > Best method, however, is to teach users proper oil application and
> ensure
> > > they clean up after themselves.
> > >
> > > Regards,
> > > Vickie
> > >
> > > Victoria Centonze Frohlich, PhD
> > > Director, Light Microscopy Shared Resource
> > > Cell & Tissue Imaging Center
> > > St. Jude Children’s Research Hospital
> > > 262 Danny Thomas Place
> > > MS 312
> > > Memphis, TN 38105
> > >
> > > Office: D1052D
> > > Phone and Digital Fax: [phone]
> > > Cell: 1-901-907-2596
> > > Paper Fax: [phone]
> > > Email: [hidden email]
> > >
> > > CTIC-LM Location: D1055 and D1056
> > > Facility Phone: [phone]
> > >
> > > http://home.stjude.org/cell-tissue-imaging/Pages/default.aspx
> > >
> > > Facility Acknowledgement for publications:
> > > “Images were acquired at the Cell & Tissue Imaging Center which is
> > > supported by SJCRH and NCI P30 CA021765.”
> > >
> > > -----Original Message-----
> > > From: Confocal Microscopy List <[hidden email]> On
> > > Behalf Of Michael Stanley
> > > Sent: Tuesday, October 27, 2020 8:19 AM
> > > To: [hidden email]
> > > Subject: Re: Protective window for a confocal microscope?
> > >
> > > Caution: External Sender
> > >
> > >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > >
> > >
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=WiaypCuZUL5s03B%2Br1rcmMLyOazUGYLj%2BmW7hRFcyIU%3D&amp;reserved=0
> > > Post images on
> > >
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=DKQAcpfaPdTDony5B7X%2Fdi%2BCVChjA7voJxvlDKq9suo%3D&amp;reserved=0
> > > and include the link in your posting.
> > > *****
> > >
> > > in the 'old days' in the core we used to cut filter paper and then
> bore a
> > > hole with a cork-bore slightly smaller than the lens.  slide 2-3 (or as
> > > many as will fit) layers of the filter paper over the objective.  this
> > will
> > > not stop a "flood" of oil (which sounds very possible for some) but it
> > will
> > > hold much more than you would think.  and, the most important feature
> of
> > > this little trick is that the oil will show very quickly on the paper
> and
> > > hopefully will trigger a response.!
> > >
> > > some of the darker filter papers will show the oil better, but this
> will
> > > vary by manufacturer.
> > >
> > > no commercial interest here, just trying to protect all the optics in
> the
> > > system.
> > >
> > > michael
> > >
> > > c. michael stanley, phd
> > > senior applications scientist
> > > chroma technology corp.
> > > 10 imtec lane
> > > bellows falls,  vt    05101
> > > [hidden email]
> > > [hidden email]
> > >
> > >
> > > -----Original Message-----
> > > From: Confocal Microscopy List <[hidden email]> On
> > > Behalf Of Gary Laevsky
> > > Sent: Tuesday, October 27, 2020 10:19 AM
> > > To: [hidden email]
> > > Subject: Re: Protective window for a confocal microscope?
> > >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > >
> > >
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=WiaypCuZUL5s03B%2Br1rcmMLyOazUGYLj%2BmW7hRFcyIU%3D&amp;reserved=0
> > > Post images on
> > >
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=yhWCT39l%2FiZJ9i8H0f4EM7MIstxliSvBVsjVQVNN8cI%3D&amp;reserved=0
> > > and include the link in your posting.
> > > *****
> > >
> > > The hair ties work great with objectives that have enough of a neck for
> > > the tie to sit on, next to the barrel.
> > >
> > > Our 40X objective (most frequently used) has no/very small neck, so the
> > > hair ties don't work.
> > >
> > > I fairly regularly (bi-weekly) have to take the objective off, store it
> > in
> > > it's case upside down, and put it in a 37C oven.  Ridiculous, in a bad
> > way,
> > > how much oil comes out.  The 40X also doesn't have "lip" that acts as a
> > bit
> > > of a reservoir (unlike the 60 and 100).
> > >
> > > On one of my heavily used systems, I have to go so far as to dismantle
> > the
> > > top (stage, objective turret, and fluorescent turret) to get to the
> piece
> > > of glass that prevents leakage into the body (amazingly awesome feature
> > of
> > > the stand).  Takes me 15 minutes.  Although I shouldn't have to do that
> > ...
> > >
> > > On Tue, Oct 27, 2020 at 9:13 AM Cammer, Michael <
> > > [hidden email]> wrote:
> > >
> > > > *****
> > > > To join, leave or search the confocal microscopy listserv, go to:
> > > >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists
> > > > .umn.edu
> %2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=04%7C01%7Cv
> > > > ictoria.frohlich%40STJUDE.ORG
> %7C6b89a1a0f4cc42f5937e08d87a8d1047%7C223
> > > >
> 40fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573459000%7CUnknown%7
> > > >
> CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXV
> > > >
> CI6Mn0%3D%7C1000&amp;sdata=q2oFBgmsbEfra%2BIBWB6i8xwEJ%2B8SF%2F59WSo3Y
> > > > %2BKvFKg%3D&amp;reserved=0 Post images on
> > > >
> > >
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=yhWCT39l%2FiZJ9i8H0f4EM7MIstxliSvBVsjVQVNN8cI%3D&amp;reserved=0
> > > and include the link in your posting.
> > > > *****
> > > >
> > > > Aquastop works well for big spills when installed correctly and has
> > > > proven effective on some of our Zeiss scopes.  But we did have a user
> > > > who spilled over or around it on our Zeiss 710, so it is not
> foolproof.
> > > >
> > > >
> > > > Hair elastics are great for the oil.
> > > >
> > > >
> > > > Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory
> > > >
> > > > NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York,
> > > > NY
> > > > 10016
> > > >
> > > > [hidden email]<mailto:[hidden email]>
> > > >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fnyulm
> > > > c.org%2Fmicros&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG
> %7C6b8
> > > >
> 9a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7
> > > >
> C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLC
> > > >
> JQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=Ud4O3pAe
> > > > 6c4HlzFgcjkQBvmAAz45ILNKOj8mnQScAsg%3D&amp;reserved=0
> > > >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fmicro
> > > > scopynotes.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG
> %7C
> > > >
> 6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C
> > > >
> 0%7C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDA
> > > >
> iLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=e6srs
> > > > AN7BHwtFBKtNbXavROHTyX2m0CdnPe3l7VnUPA%3D&amp;reserved=0
> > > >
> > > > Voice direct only, no text or messages:  1-914-309-3270 and
> > > > 1-646-501-0567
> > > >
> > > >
> > > >
> > > > ________________________________
> > > > From: Confocal Microscopy List <[hidden email]> on
> > > > behalf of Doug Richardson <[hidden email]>
> > > > Sent: Tuesday, October 27, 2020 7:41:45 AM
> > > > To: [hidden email]
> > > > Subject: Re: Protective window for a confocal microscope?
> > > >
> > > > [EXTERNAL]
> > > >
> > > > *****
> > > > To join, leave or search the confocal microscopy listserv, go to:
> > > >
> > > >
> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-
> > > >
> 2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeEl
> > > >
> Zfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=ke
> > > >
> g1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=k-eCFZaqV2J3Nbh8c16znCG7uP
> > > > Pgwni6bBeSRyqYRAE&e=
> > > > Post images on
> > > >
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=Dw
> > > >
> IFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC5
> > > >
> 0tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-d
> > > > FQ&s=o9KAzzxXBHkEPM_YiZYo9mb7m2x6GjWRoUHh-BzcNSg&e=
> > > > and include the link in your posting.
> > > > *****
> > > >
> > > > I would highly recommend the aquastop system from Zeiss. It is
> > > > overpriced for some pieces of plastic, but it forms a very tight seal
> > > > and pays for itself many times over. It is far superior to hair
> > > > elastics/glove fingers/condoms which we used in the past.
> > > >
> > > > Doug
> > > >
> > > > Get Outlook for Android
> > > >
> > > > ________________________________
> > > > From: Confocal Microscopy List <[hidden email]> on
> > > > behalf of Tomek Węgierski <[hidden email]>
> > > > Sent: Tuesday, October 27, 2020 7:12:10 AM
> > > > To: [hidden email]
> > > > <[hidden email]>
> > > > Subject: Protective window for a confocal microscope?
> > > >
> > > > *****
> > > > To join, leave or search the confocal microscopy listserv, go to:
> > > >
> > > >
> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-
> > > >
> 2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeEl
> > > >
> Zfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=ke
> > > >
> g1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=k-eCFZaqV2J3Nbh8c16znCG7uP
> > > > Pgwni6bBeSRyqYRAE&e=
> > > > Post images on
> > > >
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=Dw
> > > >
> IFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC5
> > > >
> 0tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-d
> > > > FQ&s=o9KAzzxXBHkEPM_YiZYo9mb7m2x6GjWRoUHh-BzcNSg&e=
> > > > and include the link in your posting.
> > > > *****
> > > >
> > > > Dear All,
> > > > I am considering a purchase of a protective window for our Zeiss
> > > > LSM800 and I would like to hear your advice whether this is a good
> > > > idea, and if yes, which one to buy.
> > > > The reason is our LSM800 is on an inverted stand and we already got a
> > > > leakage of oil once which went down onto Optovar. And this was
> despite
> > > > my regular checks of objectives (both sides) for leaking oil and
> > > > extensive education of users on how important it is not to use too
> > > > much oil and to clean lenses, especially on inverted stands.
> > > > Zeiss Observer has a dummy slider with an opening for 32 mm filters.
> I
> > > > hope, a protective window there should stop the oil, if the leakage
> > > > happens again.
> > > > 1) do you think placing a protective window in a confocal microscope
> > > > is a good idea or not really, because the imaging will be affected?
> > > > This would be in infinity space, so I guess it should not be
> > > > detrimental....
> > > > 2) which characteristics of protective window I should take into
> > > > consideration (substrate, thickness, coating, flatness, surface
> > > > quality, parallelism)? Currently I am inclined into protective
> glasses
> > > > by Edmund Optics such as 1-mm thick MgF2-coated Sapphire window,
> which
> > > > has high transmission, or 2-mm thick MgF2-coated B270 window, which
> > > > has a bit worse transmission but better surface parameters (and is
> > > > much cheaper).
> > > > best regards,
> > > > Tomasz
> > > >
> > > >
> > > > Tomasz Wegierski, PhD
> > > > International Institute of Molecular and Cell Biology Trojdena 4,
> > > > 02-109 Warsaw, POLAND
> > > > tel: [phone]
> > > > fax: [phone]
> > > >
> > > >
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.iimcb.gov.pl_&
> > > >
> d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDi
> > > >
> PlC50tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNB
> > > > tS-dFQ&s=u7qHptX43byvVfMe-Zn04ev5KF4-ToW1ss01pq0eVtM&e=
> > > >
> > > > ------------------------------------------------------------
> > > > This email message, including any attachments, is for the sole use of
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> > > > proprietary, confidential, and exempt from disclosure under
> applicable
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> > > > =================================
> > > >
> > >
> > >
> > > --
> > > Best,
> > >
> > > Gary Laevsky, Ph.D.
> > > Director, Confocal Imaging Facility
> > > Nikon Center of Excellence
> > > Co-Founder, North Atlantic Microscopy Society (NAMS)
> > >
> > >
> >
> https://nam11.safelinks.protection.outlook.com/?url=https%3A%2F%2Fnamsmicroscopy.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=7foXnZ0OuaU32Fj2dlCniXz%2Bn8iWTBvMyOkGj0rTX10%3D&amp;reserved=0
> > > Dept. of Molecular Biology Washington Rd.
> > > Princeton University
> > > Princeton, New Jersey, 08544-1014
> > > (O) 609 258 5432
> > > (C) 508 507 1310
> > >
> > > ________________________________
> > >
> > > Email Disclaimer: www.stjude.org/emaildisclaimer<<http://www.stjude.org/emaildisclaimer<>
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> > >
> >
> >
> > --
> > --
> > Zdenek Svindrych, Ph.D.
> > Research Scientist - Microscopy Imaging Specialist
> > Department of Biochemistry and Cell Biology
> > Geisel School of Medicine at Dartmouth
> >
>
>
> --
> Best,
>
> Gary Laevsky, Ph.D.
> Director, Confocal Imaging Facility
> Nikon Center of Excellence
> Co-Founder, North Atlantic Microscopy Society (NAMS)
> https://namsmicroscopy.com/
> Dept. of Molecular Biology
> Washington Rd.
> Princeton University
> Princeton, New Jersey, 08544-1014
> (O) 609 258 5432
> (C) 508 507 1310
>

--
--
Zdenek Svindrych, Ph.D.
Research Scientist - Microscopy Imaging Specialist
Department of Biochemistry and Cell Biology
Geisel School of Medicine at Dartmouth

[Tomek Węgierski]

*****
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Post images on http://www.imgur.com and include the link in your posting.
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Dear All,
As promised, I can give you now some feedback on the protective window:
I purchased the item #47-838 which is a 30-mm VIS-coated silica window with the parameters:
thickness 4mm, parallelism <5 arcsec, surface quality 20-10, and transmitted wavefront lambda/10.
It costs 160 EUR, but I paid 20% less due to some promotion action + transport.
1.) the opening in the filter holder is tilted, so the protective window after the installation is also tilted in the optic path,
to minimize reflections
2.) the protective filter is 30mm in diameter, which is a bit small for filter holder (32 mm), and to make it sit tight I will have to order an appropriate  O-ring. Only then the protective window should provide full protection against dripping oil. But even without the O-ring, the protective window is larger than the aperture of the filter slider, so it sits nicely in place.
3.) I checked the PSFs of 200nm beads, and I cannot tell the difference in images with and without protective window. I could not measure resolutions, because I slightly overexposed the PSFs, but PSF shapes in xy, xz, and yz planes looked the same to me.
4.) I noticed a slight increase in lateral shift between the green and far red channels: from ca. 30 nm to ca. 50 nm. This is a bit surprising to me, as there should be no bending of rays, and besides the Abbe number is not so bad (67.7). I did not check green vs red channel, because I do not have a proper prep at the moment.
In summary, I think the protective window is suitable for confocal imaging. For critical applications, it can be easily removed from the path. I feel much safer when this protective window is in the path, especially these days, when I am not so often at the institute because of COVID-19 situation.
If I discover any more disadvantages of this protective window, I will let you know.
best regards,
Tomasz

Tomasz Wegierski, PhD
International Institute of Molecular and Cell Biology
Trojdena 4, 02-109 Warsaw, POLAND
tel: [phone]
fax: [phone]
http://www.iimcb.gov.pl/
________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Tomek Węgierski <[hidden email]>
Sent: Thursday, October 29, 2020 9:05 AM
To: [hidden email] <[hidden email]>
Subject: Re: Protective window for a confocal microscope?

*****
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Dear All,
Following advice of Zdenek (thank you!) on the importance of TWE I decided to order this:
https://www.edmundoptics.de/p/30mm-dia-4mm-thick-vis-0deg-coated-lambda10-fused-silica-window/7997/
Its transmitted wavefront is lambda/10, and it shows very good parallelism, surface finish and transmission for VIS light, which is sufficient for us.
I guess the opening for filters in the filter slider is angled, but I will check this. And the slider can be removed anytime from the optic path in no time, so it will be very easy to check the imaging with and without the protective window.
When I test the PSFs I will let you know about the results.
Best,
Tomasz
PS. In my previous mail I meant 40x/1.3 EC Plan-Neofluar lens rather than "Fluar", which is a different lens (and less suitable for confocal). So, I meant one thing but somehow wrote the other. Sorry for that.

Tomasz Wegierski, PhD
International Institute of Molecular and Cell Biology
Trojdena 4, 02-109 Warsaw, POLAND
tel: [phone]
fax: [phone]
http://www.iimcb.gov.pl/
________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Zdenek Svindrych <[hidden email]>
Sent: Wednesday, October 28, 2020 12:50 PM
To: [hidden email] <[hidden email]>
Subject: Re: Protective window for a confocal microscope?

*****
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Hi,

most microscopes contain some sort of angled beam splitter or dichroic
filter in the infinity space. And, indeed, most flat surfaces (windows,
filters) are angled.

zdenek


On Tue, Oct 27, 2020 at 8:11 PM Gary Laevsky <[hidden email]>
wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi All,
>
> Staying on the question, in regards to putting a piece of glass in that
> slider (which would make our lives easier), should it be placed on an
> angle, to minimize reflections?  But, will that increase aberration?
>
> Good question Tomasz.
>
> On Tue, Oct 27, 2020 at 7:46 PM Zdenek Svindrych <[hidden email]>
> wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > Hi Tomasz,
> >
> > back to your question. The filter holder is there to hold a filter. Any
> > decent glass or fused silica AR window should do the trick.
> > As far as I remember, the hole is quite big, around 30 mm diameter
> (bigger
> > than the standard 25 mm filters), which may limit your options.
> >
> > Most filters and windows have specified surface flatness (reflected wave
> > error), but it's actually transmitted wave error (TWE) that matters here.
> > Usually, windows with single digit flatness (in lambda or fringe units)
> > will have TWE << 1/5 lambda, and should be OK.
> >
> > Thicker substrate (2 - 3 mm) usually means precision optics, and is a
> safe
> > bet. Hard coating is a must, if you want to wipe oil off of it regularly.
> > All reputable brands use hard coatings these days, 1% loss (per pass) is
> > OK.
> >
> > Sapphire is hard to work with and comes with a price premium. Fused
> silica,
> > e.g.
> >
> >
> https://www.edmundoptics.com/p/30mm-dia-vis-nir-coated-1lambda-fused-silica-window/10158/
> > (no commercial interest) looks like a good option, but BK-7 will work
> well,
> > too.
> >
> > If you happen to have a piece of AR-coated glass at hand, just put it
> there
> > and look at some beads. If you're not able to tell the difference, you're
> > good to go. You can even try a glass coverslip, the 4% loss (the
> excitation
> > loss is not an issue, you can always crank up the laser) will be hardly
> > noticeable. Coverslips are not polished, so the TWE is generally bad, but
> > with some luck you can find a good one in the stack...
> >
> > Best, zdenek
> >
> >
> > On Tue, Oct 27, 2020 at 1:18 PM Tomek Węgierski <[hidden email]>
> > wrote:
> >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > > Post images on http://www.imgur.com and include the link in your
> > posting.
> > > *****
> > >
> > > Dear All,
> > >
> > > Thank you very much for so many responses, although the discussion
> went a
> > > bit into a direction that I did not anticipate ??
> > > But yes, it is all about preventing oil from getting into the
> microscope.
> > > Our 40x/1.3 Fluar lens takes a lot of oil and I frequently have to
> clean
> > > its bottom side from oil. In contrast, I never saw any oil on the back
> of
> > > 63x/1.4 even though it is used as frequently as the 40x.
> > >
> > > I have to admit I have been skeptical about hair elastics, because when
> > > they saturate with oil they will just let it go. IMHO, it is replacing
> > "oil
> > > cleaning" approach with "oil buffering" approach. The "cleaning"
> approach
> > > has worked for us for many years. It failed when an unexperienced user
> > (who
> > > apparently did not take seriously what I was talking during the
> training)
> > > made two mistakes: did not clean the lens and let it stay in the optic
> > path
> > > after the work instead of changing for 10x. In addition, it was a
> holiday
> > > time, the system was not used for the next few days, giving the oil
> > plenty
> > > of time. So, it was a coincidence of several factors, but it can happen
> > > again.
> > >
> > > I think, hearing so many positive opinions about hair elastics, I will
> > > give them the 2nd chance in our facility. Aqua-Stop in my opinion is
> > > overpriced, and we do not currently do any experiments under perfusion
> to
> > > justify such purchase (this may change in the future). However, I would
> > not
> > > like to give up on the protective window strategy. I think placing a
> > glass
> > > element in the infinity space is not unusual. It would function as the
> > last
> > > stronghold against the oil. And it would be considerably cheaper than
> > > Aqua-Stop. I would of course test PSF. If PSF is not affected, I guess
> > the
> > > protective window is OK with imaging. So, if you still have some advice
> > > which one would be most suitable, I would appreciate it.
> > >
> > > Best,
> > > Tomasz
> > >
> > > Tomasz Wegierski, PhD
> > > International Institute of Molecular and Cell Biology
> > > Trojdena 4, 02-109 Warsaw, POLAND
> > > tel: [phone]
> > > fax: [phone]
> > > http://www.iimcb.gov.pl/
> > > ________________________________
> > > From: Confocal Microscopy List <[hidden email]> on
> > > behalf of Frohlich, Victoria <[hidden email]>
> > > Sent: Tuesday, October 27, 2020 4:38 PM
> > > To: [hidden email] <[hidden email]
> >
> > > Subject: Re: Protective window for a confocal microscope?
> > >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > > Post images on http://www.imgur.com and include the link in your
> > posting.
> > > *****
> > >
> > > Just my 2 cents.
> > > Scrunchies work well but are not fool-proof.  Cotton not polyester
> > > material are more absorbent.
> > > In addition to scrunchies I suggest wrapping the lens barrel near the
> > nose
> > > of the objective with plumbers tape to prevent oil from seeping into
> the
> > > barrel.  It can be done so that the lens retraction still functions.
> > Then
> > > once the lens is on the nosepiece of the scope the plumbers tape can be
> > > wrapped there as well.
> > >
> > > I've also used thin rubber sheets or parafilm sheets, as well as filter
> > > paper (thanks Mike S.!), to draw oil away from the lens and nosepiece.
> > >
> > > Best method, however, is to teach users proper oil application and
> ensure
> > > they clean up after themselves.
> > >
> > > Regards,
> > > Vickie
> > >
> > > Victoria Centonze Frohlich, PhD
> > > Director, Light Microscopy Shared Resource
> > > Cell & Tissue Imaging Center
> > > St. Jude Children’s Research Hospital
> > > 262 Danny Thomas Place
> > > MS 312
> > > Memphis, TN 38105
> > >
> > > Office: D1052D
> > > Phone and Digital Fax: [phone]
> > > Cell: 1-901-907-2596
> > > Paper Fax: [phone]
> > > Email: [hidden email]
> > >
> > > CTIC-LM Location: D1055 and D1056
> > > Facility Phone: [phone]
> > >
> > > http://home.stjude.org/cell-tissue-imaging/Pages/default.aspx
> > >
> > > Facility Acknowledgement for publications:
> > > “Images were acquired at the Cell & Tissue Imaging Center which is
> > > supported by SJCRH and NCI P30 CA021765.”
> > >
> > > -----Original Message-----
> > > From: Confocal Microscopy List <[hidden email]> On
> > > Behalf Of Michael Stanley
> > > Sent: Tuesday, October 27, 2020 8:19 AM
> > > To: [hidden email]
> > > Subject: Re: Protective window for a confocal microscope?
> > >
> > > Caution: External Sender
> > >
> > >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > >
> > >
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=WiaypCuZUL5s03B%2Br1rcmMLyOazUGYLj%2BmW7hRFcyIU%3D&amp;reserved=0
> > > Post images on
> > >
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=DKQAcpfaPdTDony5B7X%2Fdi%2BCVChjA7voJxvlDKq9suo%3D&amp;reserved=0
> > > and include the link in your posting.
> > > *****
> > >
> > > in the 'old days' in the core we used to cut filter paper and then
> bore a
> > > hole with a cork-bore slightly smaller than the lens.  slide 2-3 (or as
> > > many as will fit) layers of the filter paper over the objective.  this
> > will
> > > not stop a "flood" of oil (which sounds very possible for some) but it
> > will
> > > hold much more than you would think.  and, the most important feature
> of
> > > this little trick is that the oil will show very quickly on the paper
> and
> > > hopefully will trigger a response.!
> > >
> > > some of the darker filter papers will show the oil better, but this
> will
> > > vary by manufacturer.
> > >
> > > no commercial interest here, just trying to protect all the optics in
> the
> > > system.
> > >
> > > michael
> > >
> > > c. michael stanley, phd
> > > senior applications scientist
> > > chroma technology corp.
> > > 10 imtec lane
> > > bellows falls,  vt    05101
> > > [hidden email]
> > > [hidden email]
> > >
> > >
> > > -----Original Message-----
> > > From: Confocal Microscopy List <[hidden email]> On
> > > Behalf Of Gary Laevsky
> > > Sent: Tuesday, October 27, 2020 10:19 AM
> > > To: [hidden email]
> > > Subject: Re: Protective window for a confocal microscope?
> > >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > >
> > >
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=WiaypCuZUL5s03B%2Br1rcmMLyOazUGYLj%2BmW7hRFcyIU%3D&amp;reserved=0
> > > Post images on
> > >
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=yhWCT39l%2FiZJ9i8H0f4EM7MIstxliSvBVsjVQVNN8cI%3D&amp;reserved=0
> > > and include the link in your posting.
> > > *****
> > >
> > > The hair ties work great with objectives that have enough of a neck for
> > > the tie to sit on, next to the barrel.
> > >
> > > Our 40X objective (most frequently used) has no/very small neck, so the
> > > hair ties don't work.
> > >
> > > I fairly regularly (bi-weekly) have to take the objective off, store it
> > in
> > > it's case upside down, and put it in a 37C oven.  Ridiculous, in a bad
> > way,
> > > how much oil comes out.  The 40X also doesn't have "lip" that acts as a
> > bit
> > > of a reservoir (unlike the 60 and 100).
> > >
> > > On one of my heavily used systems, I have to go so far as to dismantle
> > the
> > > top (stage, objective turret, and fluorescent turret) to get to the
> piece
> > > of glass that prevents leakage into the body (amazingly awesome feature
> > of
> > > the stand).  Takes me 15 minutes.  Although I shouldn't have to do that
> > ...
> > >
> > > On Tue, Oct 27, 2020 at 9:13 AM Cammer, Michael <
> > > [hidden email]> wrote:
> > >
> > > > *****
> > > > To join, leave or search the confocal microscopy listserv, go to:
> > > >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists
> > > > .umn.edu
> %2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=04%7C01%7Cv
> > > > ictoria.frohlich%40STJUDE.ORG
> %7C6b89a1a0f4cc42f5937e08d87a8d1047%7C223
> > > >
> 40fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573459000%7CUnknown%7
> > > >
> CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXV
> > > >
> CI6Mn0%3D%7C1000&amp;sdata=q2oFBgmsbEfra%2BIBWB6i8xwEJ%2B8SF%2F59WSo3Y
> > > > %2BKvFKg%3D&amp;reserved=0 Post images on
> > > >
> > >
> >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=yhWCT39l%2FiZJ9i8H0f4EM7MIstxliSvBVsjVQVNN8cI%3D&amp;reserved=0
> > > and include the link in your posting.
> > > > *****
> > > >
> > > > Aquastop works well for big spills when installed correctly and has
> > > > proven effective on some of our Zeiss scopes.  But we did have a user
> > > > who spilled over or around it on our Zeiss 710, so it is not
> foolproof.
> > > >
> > > >
> > > > Hair elastics are great for the oil.
> > > >
> > > >
> > > > Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory
> > > >
> > > > NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York,
> > > > NY
> > > > 10016
> > > >
> > > > [hidden email]<mailto:[hidden email]>
> > > >
> https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fnyulm
> > > > c.org%2Fmicros&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG
> %7C6b8
> > > >
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> > > >
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> > > > 6c4HlzFgcjkQBvmAAz45ILNKOj8mnQScAsg%3D&amp;reserved=0
> > > >
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> > > > scopynotes.com%2F&amp;data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG
> %7C
> > > >
> 6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C
> > > >
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> > > >
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> > > > AN7BHwtFBKtNbXavROHTyX2m0CdnPe3l7VnUPA%3D&amp;reserved=0
> > > >
> > > > Voice direct only, no text or messages:  1-914-309-3270 and
> > > > 1-646-501-0567
> > > >
> > > >
> > > >
> > > > ________________________________
> > > > From: Confocal Microscopy List <[hidden email]> on
> > > > behalf of Doug Richardson <[hidden email]>
> > > > Sent: Tuesday, October 27, 2020 7:41:45 AM
> > > > To: [hidden email]
> > > > Subject: Re: Protective window for a confocal microscope?
> > > >
> > > > [EXTERNAL]
> > > >
> > > > *****
> > > > To join, leave or search the confocal microscopy listserv, go to:
> > > >
> > > >
> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-
> > > >
> 2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeEl
> > > >
> Zfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=ke
> > > >
> g1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=k-eCFZaqV2J3Nbh8c16znCG7uP
> > > > Pgwni6bBeSRyqYRAE&e=
> > > > Post images on
> > > >
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=Dw
> > > >
> IFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC5
> > > >
> 0tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-d
> > > > FQ&s=o9KAzzxXBHkEPM_YiZYo9mb7m2x6GjWRoUHh-BzcNSg&e=
> > > > and include the link in your posting.
> > > > *****
> > > >
> > > > I would highly recommend the aquastop system from Zeiss. It is
> > > > overpriced for some pieces of plastic, but it forms a very tight seal
> > > > and pays for itself many times over. It is far superior to hair
> > > > elastics/glove fingers/condoms which we used in the past.
> > > >
> > > > Doug
> > > >
> > > > Get Outlook for Android
> > > >
> > > > ________________________________
> > > > From: Confocal Microscopy List <[hidden email]> on
> > > > behalf of Tomek Węgierski <[hidden email]>
> > > > Sent: Tuesday, October 27, 2020 7:12:10 AM
> > > > To: [hidden email]
> > > > <[hidden email]>
> > > > Subject: Protective window for a confocal microscope?
> > > >
> > > > *****
> > > > To join, leave or search the confocal microscopy listserv, go to:
> > > >
> > > >
> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-
> > > >
> 2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeEl
> > > >
> Zfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=ke
> > > >
> g1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-dFQ&s=k-eCFZaqV2J3Nbh8c16znCG7uP
> > > > Pgwni6bBeSRyqYRAE&e=
> > > > Post images on
> > > >
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=Dw
> > > >
> IFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC5
> > > >
> 0tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNBtS-d
> > > > FQ&s=o9KAzzxXBHkEPM_YiZYo9mb7m2x6GjWRoUHh-BzcNSg&e=
> > > > and include the link in your posting.
> > > > *****
> > > >
> > > > Dear All,
> > > > I am considering a purchase of a protective window for our Zeiss
> > > > LSM800 and I would like to hear your advice whether this is a good
> > > > idea, and if yes, which one to buy.
> > > > The reason is our LSM800 is on an inverted stand and we already got a
> > > > leakage of oil once which went down onto Optovar. And this was
> despite
> > > > my regular checks of objectives (both sides) for leaking oil and
> > > > extensive education of users on how important it is not to use too
> > > > much oil and to clean lenses, especially on inverted stands.
> > > > Zeiss Observer has a dummy slider with an opening for 32 mm filters.
> I
> > > > hope, a protective window there should stop the oil, if the leakage
> > > > happens again.
> > > > 1) do you think placing a protective window in a confocal microscope
> > > > is a good idea or not really, because the imaging will be affected?
> > > > This would be in infinity space, so I guess it should not be
> > > > detrimental....
> > > > 2) which characteristics of protective window I should take into
> > > > consideration (substrate, thickness, coating, flatness, surface
> > > > quality, parallelism)? Currently I am inclined into protective
> glasses
> > > > by Edmund Optics such as 1-mm thick MgF2-coated Sapphire window,
> which
> > > > has high transmission, or 2-mm thick MgF2-coated B270 window, which
> > > > has a bit worse transmission but better surface parameters (and is
> > > > much cheaper).
> > > > best regards,
> > > > Tomasz
> > > >
> > > >
> > > > Tomasz Wegierski, PhD
> > > > International Institute of Molecular and Cell Biology Trojdena 4,
> > > > 02-109 Warsaw, POLAND
> > > > tel: [phone]
> > > > fax: [phone]
> > > >
> > > >
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.iimcb.gov.pl_&
> > > >
> d=DwIFBA&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDi
> > > >
> PlC50tp7rW2nBkvV7fujQf0RknE5bU&m=keg1PHlpRuZ6hHabv7CvVJ2rsWSRa_2pkARNB
> > > > tS-dFQ&s=u7qHptX43byvVfMe-Zn04ev5KF4-ToW1ss01pq0eVtM&e=
> > > >
> > > > ------------------------------------------------------------
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> > >
> > > --
> > > Best,
> > >
> > > Gary Laevsky, Ph.D.
> > > Director, Confocal Imaging Facility
> > > Nikon Center of Excellence
> > > Co-Founder, North Atlantic Microscopy Society (NAMS)
> > >
> > >
> >
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> > > Dept. of Molecular Biology Washington Rd.
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> >
> > --
> > --
> > Zdenek Svindrych, Ph.D.
> > Research Scientist - Microscopy Imaging Specialist
> > Department of Biochemistry and Cell Biology
> > Geisel School of Medicine at Dartmouth
> >
>
>
> --
> Best,
>
> Gary Laevsky, Ph.D.
> Director, Confocal Imaging Facility
> Nikon Center of Excellence
> Co-Founder, North Atlantic Microscopy Society (NAMS)
> https://namsmicroscopy.com/
> Dept. of Molecular Biology
> Washington Rd.
> Princeton University
> Princeton, New Jersey, 08544-1014
> (O) 609 258 5432
> (C) 508 507 1310
>

--
--
Zdenek Svindrych, Ph.D.
Research Scientist - Microscopy Imaging Specialist
Department of Biochemistry and Cell Biology
Geisel School of Medicine at Dartmouth

[Craig Brideau]

*****
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*****

On Mon, Nov 9, 2020 at 8:29 AM Tomek Węgierski <[hidden email]>
wrote:

> 4.) I noticed a slight increase in lateral shift between the green and far
> red channels: from ca. 30 nm to ca. 50 nm. This is a bit surprising to me,
> as there should be no bending of rays, and besides the Abbe number is not
> so bad (67.7). I did not check green vs red channel, because I do not have
> a proper prep at the moment.
>

I would expect any tilted optic in the light path to cause some lateral
chromatic aberration, even for low dispersion glass. The only way to
completely eliminate this would be to have an identical optic in the same
path tilted the opposite way. Still, 30-50nm is a rather minimal shift and
many microscopists could live with that for the sake of protecting the
internals of the microscope.

Craig