http://confocal-microscopy-list.275.s1.nabble.com/Protective-window-for-a-confocal-microscope-tp7591445p7591466.html
> *****
> 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: +48-22 597 0763
>>> fax: +48 22 597 0715
>>>
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: 1-901-595-2536
>>> Cell: 1-901-907-2596
>>> Paper Fax: 1-901-595-2909
>>> Email:
[hidden email]
>>>
>>> CTIC-LM Location: D1055 and D1056
>>> Facility Phone: 1-901-595-3439
>>>
>>>
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&data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=WiaypCuZUL5s03B%2Br1rcmMLyOazUGYLj%2BmW7hRFcyIU%3D&reserved=0>>> Post images on
>>>
>>
https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=DKQAcpfaPdTDony5B7X%2Fdi%2BCVChjA7voJxvlDKq9suo%3D&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&data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573449006%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=WiaypCuZUL5s03B%2Br1rcmMLyOazUGYLj%2BmW7hRFcyIU%3D&reserved=0>>> Post images on
>>>
>>
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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:
>>>>
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>>>> ictoria.frohlich%40STJUDE.ORG%7C6b89a1a0f4cc42f5937e08d87a8d1047%7C223
>>>> 40fa892264871b677d3b3e377af72%7C0%7C0%7C637394093573459000%7CUnknown%7
>>>> CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXV
>>>> CI6Mn0%3D%7C1000&sdata=q2oFBgmsbEfra%2BIBWB6i8xwEJ%2B8SF%2F59WSo3Y
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>>
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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]>
>>>>
https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fnyulm>>>> c.org%2Fmicros&data=04%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7C6b8
>>>> 9a1a0f4cc42f5937e08d87a8d1047%7C22340fa892264871b677d3b3e377af72%7C0%7
>>>> C0%7C637394093573459000%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLC
>>>> JQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=Ud4O3pAe
>>>> 6c4HlzFgcjkQBvmAAz45ILNKOj8mnQScAsg%3D&reserved=0
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>>>> iLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=e6srs
>>>> AN7BHwtFBKtNbXavROHTyX2m0CdnPe3l7VnUPA%3D&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: +48-22 597 0763
>>>> fax: +48 22 597 0715
>>>>
>>>>
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
>>
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