imaging through quartz or fused silica

[Romin, Yevgeniy/Sloan Kettering Institute]

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Dear List

Has anybody here done confocal imaging through quartz or fused silica, rather then through a glass coverslip?  One of our users is irradiating his yeast cells using UV light through the bottom of a multi well plate before doing a timelapse, but it seems that the irradiation is not at all efficient through glass.  It's recommended that it be done through quartz or fused silica.  He wants to do live imaging immediately after irradiation, so he does not have the time to transfer the yeast anywhere, he has to image in whatever he irradiated in.  He can have a plate custom made (with quartz cut to glass coverslip thickness), and I was wondering if anybody had any experience here with something similar.

Thanks to all of you in advance,

Yevgeniy Romin
 
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[John Oreopoulos]

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Common glasses certainly absorb and do not transmit very well UV wavelengths. As I recall Olympus manufactured some time ago a TIRF objective with an NA of 1.65 and subsequently required that the sample be mounted on quartz coverslips and imaged with an immersion oil with a very high refractive index (no commercial interest). We had one of these objectives in the lab I worked in during my PhD, but I never used it. As I recall, there was a large bag of quartz coverslips on hand to be used with this objective, and I believe the coverslips were 1" round diameter made by SPI (no commercial interest):

http://www.2spi.com/catalog/ltmic/quartz.shtml

These coverslips can be mounted in a standard coverslip holder, like the one that Invitrogen makes (no commercial interest):

http://products.invitrogen.com/ivgn/product/A7816

Maybe if you ordered a few chambers like these it could do the same job as a custom multi-well plate. Might be a bit cheaper too in the long run. Make sure your objective also has good UV transmission before trying it this way.


John Oreopoulos
Research Assistant
Spectral Applied Research
Richmond Hill, Ontario
Canada
www.spectral.ca



On 2011-12-21, at 5:47 PM, Yevgeniy Romin wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear List
>
> Has anybody here done confocal imaging through quartz or fused silica, rather then through a glass coverslip?  One of our users is irradiating his yeast cells using UV light through the bottom of a multi well plate before doing a timelapse, but it seems that the irradiation is not at all efficient through glass.  It's recommended that it be done through quartz or fused silica.  He wants to do live imaging immediately after irradiation, so he does not have the time to transfer the yeast anywhere, he has to image in whatever he irradiated in.  He can have a plate custom made (with quartz cut to glass coverslip thickness), and I was wondering if anybody had any experience here with something similar.
>
> Thanks to all of you in advance,
>
> Yevgeniy Romin
>
>     =====================================================================
>
>     Please note that this e-mail and any files transmitted with it may be
>     privileged, confidential, and protected from disclosure under
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>     recipient, or an employee or agent responsible for delivering this
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>     computer.

[mcammer]

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In a previous lab we had one of these 100X Olympus objectives too.  We rarely used it.  For one thing, I found the immersion medium very irritating to my skin and it was really too thin to use with an inverted microscope.  But this discussion is about the coverslips.

The coverslips were sapphire and very expensive.  They were very brittle.  Therefore, it was very difficult to use them.  I don't know about the UV transmission properties of the sapphire.  

I think there was a discussion about quartz coverslips in the past year or two (ot three?) on this listserv or the microscopy one.  Recommend doing searches of these repositories of knowledge.
_________________________________________
Michael Cammer, Assistant Research Scientist
Skirball Institute of Biomolecular Medicine
Lab: [phone]  Cell: [phone]

________________________________________
From: Confocal Microscopy List [[hidden email]] On Behalf Of John Oreopoulos [[hidden email]]
Sent: Wednesday, December 21, 2011 6:05 PM
To: [hidden email]
Subject: Re: imaging through quartz or fused silica

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Common glasses certainly absorb and do not transmit very well UV wavelengths. As I recall Olympus manufactured some time ago a TIRF objective with an NA of 1.65 and subsequently required that the sample be mounted on quartz coverslips and imaged with an immersion oil with a very high refractive index (no commercial interest). We had one of these objectives in the lab I worked in during my PhD, but I never used it. As I recall, there was a large bag of quartz coverslips on hand to be used with this objective, and I believe the coverslips were 1" round diameter made by SPI (no commercial interest):

http://www.2spi.com/catalog/ltmic/quartz.shtml

These coverslips can be mounted in a standard coverslip holder, like the one that Invitrogen makes (no commercial interest):

http://products.invitrogen.com/ivgn/product/A7816

Maybe if you ordered a few chambers like these it could do the same job as a custom multi-well plate. Might be a bit cheaper too in the long run. Make sure your objective also has good UV transmission before trying it this way.


John Oreopoulos
Research Assistant
Spectral Applied Research
Richmond Hill, Ontario
Canada
www.spectral.ca



On 2011-12-21, at 5:47 PM, Yevgeniy Romin wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear List
>
> Has anybody here done confocal imaging through quartz or fused silica, rather then through a glass coverslip?  One of our users is irradiating his yeast cells using UV light through the bottom of a multi well plate before doing a timelapse, but it seems that the irradiation is not at all efficient through glass.  It's recommended that it be done through quartz or fused silica.  He wants to do live imaging immediately after irradiation, so he does not have the time to transfer the yeast anywhere, he has to image in whatever he irradiated in.  He can have a plate custom made (with quartz cut to glass coverslip thickness), and I was wondering if anybody had any experience here with something similar.
>
> Thanks to all of you in advance,
>
> Yevgeniy Romin
>
>     =====================================================================
>
>     Please note that this e-mail and any files transmitted with it may be
>     privileged, confidential, and protected from disclosure under
>     applicable law. If the reader of this message is not the intended
>     recipient, or an employee or agent responsible for delivering this
>     message to the intended recipient, you are hereby notified that any
>     reading, dissemination, distribution, copying, or other use of this
>     communication or any of its attachments is strictly prohibited.  If
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>     sender immediately by replying to this message and deleting this
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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.
=================================

[John Oreopoulos]

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Yes, you're right Michael, the coverslips were saphire, not quartz. I was mistaken. Saphire has an index of refraction that makes it compatible with the high NA objective. Anyways, as you say, the discussion is about quartz coverslips, and there was another conversation about this fairly recently I think.

John Oreopoulos

On 2011-12-21, at 8:42 PM, "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
> *****
>
> In a previous lab we had one of these 100X Olympus objectives too.  We rarely used it.  For one thing, I found the immersion medium very irritating to my skin and it was really too thin to use with an inverted microscope.  But this discussion is about the coverslips.
>
> The coverslips were sapphire and very expensive.  They were very brittle.  Therefore, it was very difficult to use them.  I don't know about the UV transmission properties of the sapphire.  
>
> I think there was a discussion about quartz coverslips in the past year or two (ot three?) on this listserv or the microscopy one.  Recommend doing searches of these repositories of knowledge.
> _________________________________________
> Michael Cammer, Assistant Research Scientist
> Skirball Institute of Biomolecular Medicine
> Lab: [phone]  Cell: [phone]
>
> ________________________________________
> From: Confocal Microscopy List [[hidden email]] On Behalf Of John Oreopoulos [[hidden email]]
> Sent: Wednesday, December 21, 2011 6:05 PM
> To: [hidden email]
> Subject: Re: imaging through quartz or fused silica
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Common glasses certainly absorb and do not transmit very well UV wavelengths. As I recall Olympus manufactured some time ago a TIRF objective with an NA of 1.65 and subsequently required that the sample be mounted on quartz coverslips and imaged with an immersion oil with a very high refractive index (no commercial interest). We had one of these objectives in the lab I worked in during my PhD, but I never used it. As I recall, there was a large bag of quartz coverslips on hand to be used with this objective, and I believe the coverslips were 1" round diameter made by SPI (no commercial interest):
>
> http://www.2spi.com/catalog/ltmic/quartz.shtml
>
> These coverslips can be mounted in a standard coverslip holder, like the one that Invitrogen makes (no commercial interest):
>
> http://products.invitrogen.com/ivgn/product/A7816
>
> Maybe if you ordered a few chambers like these it could do the same job as a custom multi-well plate. Might be a bit cheaper too in the long run. Make sure your objective also has good UV transmission before trying it this way.
>
>
> John Oreopoulos
> Research Assistant
> Spectral Applied Research
> Richmond Hill, Ontario
> Canada
> www.spectral.ca
>
>
>
> On 2011-12-21, at 5:47 PM, Yevgeniy Romin wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Dear List
>>
>> Has anybody here done confocal imaging through quartz or fused silica, rather then through a glass coverslip?  One of our users is irradiating his yeast cells using UV light through the bottom of a multi well plate before doing a timelapse, but it seems that the irradiation is not at all efficient through glass.  It's recommended that it be done through quartz or fused silica.  He wants to do live imaging immediately after irradiation, so he does not have the time to transfer the yeast anywhere, he has to image in whatever he irradiated in.  He can have a plate custom made (with quartz cut to glass coverslip thickness), and I was wondering if anybody had any experience here with something similar.
>>
>> Thanks to all of you in advance,
>>
>> Yevgeniy Romin
>>
>>    =====================================================================
>>
>>    Please note that this e-mail and any files transmitted with it may be
>>    privileged, confidential, and protected from disclosure under
>>    applicable law. If the reader of this message is not the intended
>>    recipient, or an employee or agent responsible for delivering this
>>    message to the intended recipient, you are hereby notified that any
>>    reading, dissemination, distribution, copying, or other use of this
>>    communication or any of its attachments is strictly prohibited.  If
>>    you have received this communication in error, please notify the
>>    sender immediately by replying to this message and deleting this
>>    message, any attachments, and all copies and backups from your
>>    computer.
>
> ------------------------------------------------------------
> 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.
> =================================

[Peter Gabriel Pitrone]

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Hello Yevgeniy,

I would recommend figuring out the correct thickness of the needed
coverslips before purchasing them... If the R.I. is significantly higher
in quartz, which I believe it is (~1.6ish), then you need to calculate how
thick it needs to be to match 0.1700 mm of the type of glass used in
normal coverslips (~1.52ish).

Good luck!

Pete

On Wed, December 21, 2011 11:47 pm, Yevgeniy Romin wrote:
| *****
| To join, leave or search the confocal microscopy listserv, go to:
| http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
| *****
|
| Dear List
|
| Has anybody here done confocal imaging through quartz or fused silica,
| rather then through a glass coverslip?  One of our users is irradiating
| his yeast cells using UV light through the bottom of a multi well plate
| before doing a timelapse, but it seems that the irradiation is not at all
| efficient through glass.  It's recommended that it be done through quartz
| or fused silica.  He wants to do live imaging immediately after
| irradiation, so he does not have the time to transfer the yeast anywhere,
| he has to image in whatever he irradiated in.  He can have a plate custom
| made (with quartz cut to glass coverslip thickness), and I was wondering
| if anybody had any experience here with something similar.
|
| Thanks to all of you in advance,
|
| Yevgeniy Romin
|
|      =====================================================================
|
|      Please note that this e-mail and any files transmitted with it may be
|      privileged, confidential, and protected from disclosure under
|      applicable law. If the reader of this message is not the intended
|      recipient, or an employee or agent responsible for delivering this
|      message to the intended recipient, you are hereby notified that any
|      reading, dissemination, distribution, copying, or other use of this
|      communication or any of its attachments is strictly prohibited.  If
|      you have received this communication in error, please notify the
|      sender immediately by replying to this message and deleting this
|      message, any attachments, and all copies and backups from your
|      computer.
|


--
Peter Gabriel Pitrone - TechRMS
Microscopy/Imaging Specialist
Prof. Dr. Pavel Tomancak group
Max Planck Institute for
Molecular Biology and Genetics
Pfotenhauerstr. 108
01307 Dresden

"If a straight line fit is required, obtain only two data points." - Anon.

[Romin, Yevgeniy/Sloan Kettering Institute]

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Thanks to everybody for your advice.  I will take all this into consideration.

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Peter Gabriel Pitrone
Sent: Thursday, December 22, 2011 3:00 AM
To: [hidden email]
Subject: Re: imaging through quartz or fused silica

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Hello Yevgeniy,

I would recommend figuring out the correct thickness of the needed coverslips before purchasing them... If the R.I. is significantly higher in quartz, which I believe it is (~1.6ish), then you need to calculate how thick it needs to be to match 0.1700 mm of the type of glass used in normal coverslips (~1.52ish).

Good luck!

Pete

On Wed, December 21, 2011 11:47 pm, Yevgeniy Romin wrote:
| *****
| To join, leave or search the confocal microscopy listserv, go to:
| http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
| *****
|
| Dear List
|
| Has anybody here done confocal imaging through quartz or fused silica,
| rather then through a glass coverslip?  One of our users is
| irradiating his yeast cells using UV light through the bottom of a
| multi well plate before doing a timelapse, but it seems that the
| irradiation is not at
all
| efficient through glass.  It's recommended that it be done through
quartz
| or fused silica.  He wants to do live imaging immediately after
| irradiation, so he does not have the time to transfer the yeast
anywhere,
| he has to image in whatever he irradiated in.  He can have a plate
custom
| made (with quartz cut to glass coverslip thickness), and I was
| wondering if anybody had any experience here with something similar.
|
| Thanks to all of you in advance,
|
| Yevgeniy Romin
|
|
=====================================================================
|
|      Please note that this e-mail and any files transmitted with it
| may
be
|      privileged, confidential, and protected from disclosure under
|      applicable law. If the reader of this message is not the intended
|      recipient, or an employee or agent responsible for delivering this
|      message to the intended recipient, you are hereby notified that any
|      reading, dissemination, distribution, copying, or other use of this
|      communication or any of its attachments is strictly prohibited.  If
|      you have received this communication in error, please notify the
|      sender immediately by replying to this message and deleting this
|      message, any attachments, and all copies and backups from your
|      computer.
|


--
Peter Gabriel Pitrone - TechRMS
Microscopy/Imaging Specialist
Prof. Dr. Pavel Tomancak group
Max Planck Institute for
Molecular Biology and Genetics
Pfotenhauerstr. 108
01307 Dresden

"If a straight line fit is required, obtain only two data points." - Anon.

[Arvydas Matiukas]

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Hello list,
 
I have a question to Core Managers/Directors
regarding the maximum load (hrs/year) that a
single confocal can efficiently handle. Getting
above this limit would mean a necessity to buy
an extra confocal.
 
Our LSM510 is currently being used 1400 hrs/year
and the scheduling becomes quite problematic.
I assume the usage of 1500 hrs/year or 30hrs/week
for a 7 year old confocal is close to the physical limit
taking into account downtime (~3 weeks) and scheduling
conflicts between 10 major users.
 
Happy Holidays,
Arvydas
-------------------------------


 
 
 
Arvydas Matiukas, Ph.D.
Director of Confocal&Two-Photon Core
Department of Pharmacology
SUNY Upstate Medical University
766 Irving Ave., WH 3167
Syracuse, NY 13210
tel.: [phone]
fax: [phone]
email: [hidden email]

[George McNamara]

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8784 hours in 2012 (24 hours * 366 days), ok maybe minus ~504 hours
according to your maintenance downtime (seems excessive unless the NY
ran out of money for salt and snow plows). If they are major users, give
them 24/7 access. If your users don't like weekends or late night/early
mornings,  3,000 hours (12 hours * 250 days). I doubt you have many
scheduling conflicts at 7am, and only a few more at 7pm. If they want to
thaw out, I have hours available on the Leica SP5 inverted system with
motorized stage and MP/SP5 upright (the latter also has B&H 2 channel
TCSPC FLIM and ISS FCS ... discount if you or they can teach me FCS or
send me a review copy of a future best selling "FCS for Dummies") -
there is a hotel one block away but I suggest www.oceanfive.com or other
South Beach hotel.

Use an online scheduler - ours is a hand me down from Dana Farber (free
= you get what you pay for) that works ok - see
http://sccc.ccs.miami.edu/AICFScheduler

You can buy my core's LSM510 and ship it up state (the Fujitsu Siemens
PC died a week ago, dealing with that can be discounted in the pricing).
This is a 4 PMT system, visible Argon ion, 543 HeNe, 633 HeNe (UV laser
ready but I recommend a bottle of To-Pro-3 instead). Contact me at
[hidden email]

I recently helped a neighbor obtain quotes for $250K confocal scopes.
You can get a nice confocal for $250K (if you manage a buy one, get one
free, please send one to me). I was also impressed by the API
DeltaVision Elite system - could add the 150x/1.4? lens and 25x/0.9 4 mm
working distance objective lens, and some nice computer improvements for
under $250K.

Of course if you/your users want to do molecule scale colocalization
with Mander's, Pearson, et al - don't bother with a confocal or those
coefficients - get yourself a nanoscope (and please get me one too).

George
p.s. if you send anyone down and they bring fresh Dinosaur Bar-B-Que,
first four hours are no charge.

On 12/22/2011 6:04 PM, Arvydas Matiukas wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hello list,
>
> I have a question to Core Managers/Directors
> regarding the maximum load (hrs/year) that a
> single confocal can efficiently handle. Getting
> above this limit would mean a necessity to buy
> an extra confocal.
>
> Our LSM510 is currently being used 1400 hrs/year
> and the scheduling becomes quite problematic.
> I assume the usage of 1500 hrs/year or 30hrs/week
> for a 7 year old confocal is close to the physical limit
> taking into account downtime (~3 weeks) and scheduling
> conflicts between 10 major users.
>
> Happy Holidays,
> Arvydas
> -------------------------------
>
>
>
>
>
> Arvydas Matiukas, Ph.D.
> Director of Confocal&Two-Photon Core
> Department of Pharmacology
> SUNY Upstate Medical University
> 766 Irving Ave., WH 3167
> Syracuse, NY 13210
> tel.: [phone]
> fax: [phone]
> email: [hidden email]
>
>    

--


George McNamara, PhD
Analytical Imaging Core Facility
University of Miami

[Johannes Helm]

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Good afternoon,

I am sorry that I am late with adding a comment, the content of which
possibly already had been mentioned (although I did not find it).

While fused silica essentially is "glassy" in its character - i.e.
optically isotropic - quartz and sapphire are crystals (trigonal) with
bi-refringence properties. There will, hence, not be a single refractive
index but there will be different refractive indices for different
orientations.

I assume that the producers of quartz or sapphire cover slips take this
into account. However, when using lenses with large NAs it will be
un-avoidable to have different parts of the light cone passing the cover
slip to experience different refractive indices.



Best wishes,

Johannes

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Thanks to everybody for your advice.  I will take all this into
> consideration.
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of Peter Gabriel Pitrone
> Sent: Thursday, December 22, 2011 3:00 AM
> To: [hidden email]
> Subject: Re: imaging through quartz or fused silica
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hello Yevgeniy,
>
> I would recommend figuring out the correct thickness of the needed
> coverslips before purchasing them... If the R.I. is significantly higher
> in quartz, which I believe it is (~1.6ish), then you need to calculate how
> thick it needs to be to match 0.1700 mm of the type of glass used in
> normal coverslips (~1.52ish).
>
> Good luck!
>
> Pete
>
> On Wed, December 21, 2011 11:47 pm, Yevgeniy Romin wrote:
> | *****
> | To join, leave or search the confocal microscopy listserv, go to:
> | http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> | *****
> |
> | Dear List
> |
> | Has anybody here done confocal imaging through quartz or fused silica,
> | rather then through a glass coverslip?  One of our users is
> | irradiating his yeast cells using UV light through the bottom of a
> | multi well plate before doing a timelapse, but it seems that the
> | irradiation is not at
> all
> | efficient through glass.  It's recommended that it be done through
> quartz
> | or fused silica.  He wants to do live imaging immediately after
> | irradiation, so he does not have the time to transfer the yeast
> anywhere,
> | he has to image in whatever he irradiated in.  He can have a plate
> custom
> | made (with quartz cut to glass coverslip thickness), and I was
> | wondering if anybody had any experience here with something similar.
> |
> | Thanks to all of you in advance,
> |
> | Yevgeniy Romin
> |
> |
> =====================================================================
> |
> |      Please note that this e-mail and any files transmitted with it
> | may
> be
> |      privileged, confidential, and protected from disclosure under
> |      applicable law. If the reader of this message is not the intended
> |      recipient, or an employee or agent responsible for delivering this
> |      message to the intended recipient, you are hereby notified that any
> |      reading, dissemination, distribution, copying, or other use of this
> |      communication or any of its attachments is strictly prohibited.  If
> |      you have received this communication in error, please notify the
> |      sender immediately by replying to this message and deleting this
> |      message, any attachments, and all copies and backups from your
> |      computer.
> |
>
>
> --
> Peter Gabriel Pitrone - TechRMS
> Microscopy/Imaging Specialist
> Prof. Dr. Pavel Tomancak group
> Max Planck Institute for
> Molecular Biology and Genetics
> Pfotenhauerstr. 108
> 01307 Dresden
>
> "If a straight line fit is required, obtain only two data points." - Anon.
>

--
P. Johannes Helm

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[Craig Brideau]

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Sapphire's anisotropy is fairly weak, and its hardness makes it a somewhat
popular optical window.  You'd want the crystal axis oriented in the 'Z'
direction and even still it would slightly alter polarization along the
optical axis.  That said, since it is a relatively weak effect Sapphire
might still work for you depending on how high your NA was.

Craig


On Sat, Dec 24, 2011 at 4:39 AM, Johannes Helm <[hidden email]>wrote:

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> *****
>
> Good afternoon,
>
> I am sorry that I am late with adding a comment, the content of which
> possibly already had been mentioned (although I did not find it).
>
> While fused silica essentially is "glassy" in its character - i.e.
> optically isotropic - quartz and sapphire are crystals (trigonal) with
> bi-refringence properties. There will, hence, not be a single refractive
> index but there will be different refractive indices for different
> orientations.
>
> I assume that the producers of quartz or sapphire cover slips take this
> into account. However, when using lenses with large NAs it will be
> un-avoidable to have different parts of the light cone passing the cover
> slip to experience different refractive indices.
>
>
>
> Best wishes,
>
> Johannes
>
> > *****
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> >
> > Thanks to everybody for your advice.  I will take all this into
> > consideration.
> >
> > -----Original Message-----
> > From: Confocal Microscopy List [mailto:[hidden email]]
> > On Behalf Of Peter Gabriel Pitrone
> > Sent: Thursday, December 22, 2011 3:00 AM
> > To: [hidden email]
> > Subject: Re: imaging through quartz or fused silica
> >
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > Hello Yevgeniy,
> >
> > I would recommend figuring out the correct thickness of the needed
> > coverslips before purchasing them... If the R.I. is significantly higher
> > in quartz, which I believe it is (~1.6ish), then you need to calculate
> how
> > thick it needs to be to match 0.1700 mm of the type of glass used in
> > normal coverslips (~1.52ish).
> >
> > Good luck!
> >
> > Pete
> >
> > On Wed, December 21, 2011 11:47 pm, Yevgeniy Romin wrote:
> > | *****
> > | To join, leave or search the confocal microscopy listserv, go to:
> > | http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
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> > |
> > | Dear List
> > |
> > | Has anybody here done confocal imaging through quartz or fused silica,
> > | rather then through a glass coverslip?  One of our users is
> > | irradiating his yeast cells using UV light through the bottom of a
> > | multi well plate before doing a timelapse, but it seems that the
> > | irradiation is not at
> > all
> > | efficient through glass.  It's recommended that it be done through
> > quartz
> > | or fused silica.  He wants to do live imaging immediately after
> > | irradiation, so he does not have the time to transfer the yeast
> > anywhere,
> > | he has to image in whatever he irradiated in.  He can have a plate
> > custom
> > | made (with quartz cut to glass coverslip thickness), and I was
> > | wondering if anybody had any experience here with something similar.
> > |
> > | Thanks to all of you in advance,
> > |
> > | Yevgeniy Romin
> > |
> > |
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> >
> > --
> > Peter Gabriel Pitrone - TechRMS
> > Microscopy/Imaging Specialist
> > Prof. Dr. Pavel Tomancak group
> > Max Planck Institute for
> > Molecular Biology and Genetics
> > Pfotenhauerstr. 108
> > 01307 Dresden
> >
> > "If a straight line fit is required, obtain only two data points." -
> Anon.
> >
>
>
> --
> P. Johannes Helm
>
> Voice:  ([phone] (office)
> Fax:    [phone] (office)
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[Paul Rigby-2]

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Hi Yevgeniy,
While I have not had any experience with quartz coverslips, perhaps you might be able to achieve a better (or cheaper) result by using an upright microscope (or objective inverter) and a UV transmitting dipping objective.
Hope this might help.
Cheers
Paul

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Yevgeniy Romin
Sent: Thursday, 22 December 2011 6:47 AM
To: [hidden email]
Subject: imaging through quartz or fused silica

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Dear List

Has anybody here done confocal imaging through quartz or fused silica, rather then through a glass coverslip?  One of our users is irradiating his yeast cells using UV light through the bottom of a multi well plate before doing a timelapse, but it seems that the irradiation is not at all efficient through glass.  It's recommended that it be done through quartz or fused silica.  He wants to do live imaging immediately after irradiation, so he does not have the time to transfer the yeast anywhere, he has to image in whatever he irradiated in.  He can have a plate custom made (with quartz cut to glass coverslip thickness), and I was wondering if anybody had any experience here with something similar.

Thanks to all of you in advance,

Yevgeniy Romin