Ariel, Pablo |
*****
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 demoing a long working distance water objective from Leica (HC FLUOTAR L 25x/0,95 W VISIR, WD: 2.4mm #506375) in an inverted configuration, and am running into some challenges. While the objective is very nice, the long working distance presents a problem for long-duration timelapses with multiple positions. Water works fine, but it eventually evaporates (in hours). Zeiss Immersol W does not evaporate, but cannot hold the column when the objective is over a mm from the coverslip (it drips/slides off). Leica has an automated water dispenser, but it is not available for my current demo. I was wondering if anyone had any ideas for other immersion options that have a surface tension and RI similar to water, but don't evaporate as easily. Thanks, Pablo Pablo Ariel, Ph.D. Assistant Professor Director of the Microscopy Services Laboratory Brinkhous-Bullitt Bldg B04 Department of Pathology and Laboratory Medicine University of North Carolina at Chapel Hill http://www.med.unc.edu/microscopy Tel: 919-966-2413 |
Tilo Moede |
*****
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. ***** We use viscotears(artificial tear-fluid replacement) as immersion fluid with exactly that lens (and it's 10x and 20x relatives) for in vivo imaging of organoids transplanted to the anterior chamber of the eye with great results.. Something like that should be available from a pharmacy. Cheerio Tilo Hämta Outlook för Android<https://aka.ms/AAb9ysg> ________________________________ From: Confocal Microscopy List <[hidden email]> on behalf of Ariel, Pablo <[hidden email]> Sent: Thursday, April 15, 2021 12:05:25 PM To: [hidden email] <[hidden email]> Subject: immersion fluid options for long timelapses with long working distance water objective ***** To join, leave or search the confocal microscopy listserv, go to: https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7CTilo.Moede%40KI.SE%7C3517a286bb174a78639b08d8fff65b64%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637540780863329546%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=2qHml3iWWWFliCmDgStyuaYKjK7jnpvZhSg6vzZxSs8%3D&reserved=0 Post images on https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7CTilo.Moede%40KI.SE%7C3517a286bb174a78639b08d8fff65b64%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637540780863329546%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=rFgQ0fCjIpE%2FNn0EBPQErmRAEpYW6vQ%2BqH37KgH%2FUT0%3D&reserved=0 and include the link in your posting. ***** Dear all, I am demoing a long working distance water objective from Leica (HC FLUOTAR L 25x/0,95 W VISIR, WD: 2.4mm #506375) in an inverted configuration, and am running into some challenges. While the objective is very nice, the long working distance presents a problem for long-duration timelapses with multiple positions. Water works fine, but it eventually evaporates (in hours). Zeiss Immersol W does not evaporate, but cannot hold the column when the objective is over a mm from the coverslip (it drips/slides off). Leica has an automated water dispenser, but it is not available for my current demo. I was wondering if anyone had any ideas for other immersion options that have a surface tension and RI similar to water, but don't evaporate as easily. Thanks, Pablo Pablo Ariel, Ph.D. Assistant Professor Director of the Microscopy Services Laboratory Brinkhous-Bullitt Bldg B04 Department of Pathology and Laboratory Medicine University of North Carolina at Chapel Hill https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.med.unc.edu%2Fmicroscopy&data=04%7C01%7CTilo.Moede%40KI.SE%7C3517a286bb174a78639b08d8fff65b64%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637540780863329546%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=4wo53%2Bh3lqdhhPlnEhu2oibKqKHBI3E2kO%2BBpX4wmfo%3D&reserved=0 Tel: 919-966-2413 När du skickar e-post till Karolinska Institutet (KI) innebär detta att KI kommer att behandla dina personuppgifter. Här finns information om hur KI behandlar personuppgifter<https://ki.se/medarbetare/integritetsskyddspolicy>. Sending email to Karolinska Institutet (KI) will result in KI processing your personal data. You can read more about KI's processing of personal data here<https://ki.se/en/staff/data-protection-policy>. |
Gary Laevsky |
In reply to this post by Ariel, Pablo
*****
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. ***** Many years ago we used Suave hair gel. It worked for multiple positions fairly close together. Best, Gary > On Apr 15, 2021, at 6:08 AM, Ariel, Pablo <[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, > > I am demoing a long working distance water objective from Leica (HC FLUOTAR L 25x/0,95 W VISIR, WD: 2.4mm #506375) in an inverted configuration, and am running into some challenges. > > While the objective is very nice, the long working distance presents a problem for long-duration timelapses with multiple positions. Water works fine, but it eventually evaporates (in hours). Zeiss Immersol W does not evaporate, but cannot hold the column when the objective is over a mm from the coverslip (it drips/slides off). Leica has an automated water dispenser, but it is not available for my current demo. I was wondering if anyone had any ideas for other immersion options that have a surface tension and RI similar to water, but don't evaporate as easily. > > Thanks, > Pablo > > Pablo Ariel, Ph.D. > Assistant Professor > Director of the Microscopy Services Laboratory > Brinkhous-Bullitt Bldg B04 > Department of Pathology and Laboratory Medicine > University of North Carolina at Chapel Hill > http://www.med.unc.edu/microscopy > Tel: 919-966-2413 |
Ariel, Pablo |
In reply to this post by Tilo Moede
*****
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. ***** Thanks for the suggestion Tilo. How long do the viscotears last before evaporating? We are hoping for multiple position imaging over 48 hours. Best, Pablo -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Tilo Moede Sent: Thursday, April 15, 2021 6:15 AM To: [hidden email] Subject: Re: immersion fluid options for long timelapses with long working distance water objective ***** 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. ***** We use viscotears(artificial tear-fluid replacement) as immersion fluid with exactly that lens (and it's 10x and 20x relatives) for in vivo imaging of organoids transplanted to the anterior chamber of the eye with great results.. Something like that should be available from a pharmacy. Cheerio Tilo Hämta Outlook för Android<https://aka.ms/AAb9ysg> ________________________________ From: Confocal Microscopy List <[hidden email]> on behalf of Ariel, Pablo <[hidden email]> Sent: Thursday, April 15, 2021 12:05:25 PM To: [hidden email] <[hidden email]> Subject: immersion fluid options for long timelapses with long working distance water objective ***** To join, leave or search the confocal microscopy listserv, go to: https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7CTilo.Moede%40KI.SE%7C3517a286bb174a78639b08d8fff65b64%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637540780863329546%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=2qHml3iWWWFliCmDgStyuaYKjK7jnpvZhSg6vzZxSs8%3D&reserved=0 Post images on https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7CTilo.Moede%40KI.SE%7C3517a286bb174a78639b08d8fff65b64%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637540780863329546%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=rFgQ0fCjIpE%2FNn0EBPQErmRAEpYW6vQ%2BqH37KgH%2FUT0%3D&reserved=0 and include the link in your posting. ***** Dear all, I am demoing a long working distance water objective from Leica (HC FLUOTAR L 25x/0,95 W VISIR, WD: 2.4mm #506375) in an inverted configuration, and am running into some challenges. While the objective is very nice, the long working distance presents a problem for long-duration timelapses with multiple positions. Water works fine, but it eventually evaporates (in hours). Zeiss Immersol W does not evaporate, but cannot hold the column when the objective is over a mm from the coverslip (it drips/slides off). Leica has an automated water dispenser, but it is not available for my current demo. I was wondering if anyone had any ideas for other immersion options that have a surface tension and RI similar to water, but don't evaporate as easily. Thanks, Pablo Pablo Ariel, Ph.D. Assistant Professor Director of the Microscopy Services Laboratory Brinkhous-Bullitt Bldg B04 Department of Pathology and Laboratory Medicine University of North Carolina at Chapel Hill https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.med.unc.edu%2Fmicroscopy&data=04%7C01%7CTilo.Moede%40KI.SE%7C3517a286bb174a78639b08d8fff65b64%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637540780863329546%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=4wo53%2Bh3lqdhhPlnEhu2oibKqKHBI3E2kO%2BBpX4wmfo%3D&reserved=0 Tel: 919-966-2413 När du skickar e-post till Karolinska Institutet (KI) innebär detta att KI kommer att behandla dina personuppgifter. Här finns information om hur KI behandlar personuppgifter<https://ki.se/medarbetare/integritetsskyddspolicy>. Sending email to Karolinska Institutet (KI) will result in KI processing your personal data. You can read more about KI's processing of personal data here<https://ki.se/en/staff/data-protection-policy>. |
Cammer, Michael-2 |
*****
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. ***** A few years ago people on this listserv suggested jelly for sonograms and Genteal eye gel. Here is info on the latter http://microscopynotes.com/710/genteal/index.html 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 Ariel, Pablo <[hidden email]> Sent: Thursday, April 15, 2021 6:41:53 AM To: [hidden email] Subject: Re: immersion fluid options for long timelapses with long working distance water objective [EXTERNAL] ***** To join, leave or search the confocal microscopy listserv, go to: https://urldefense.com/v3/__http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy__;!!MXfaZl3l!NV0LInRfEiVhwJn3wg12JwYCTat4uHR7sWvEWVnmG_jiEDgNogdg1LQn_Un-qnVTiX8EklI$ Post images on https://urldefense.com/v3/__http://www.imgur.com__;!!MXfaZl3l!NV0LInRfEiVhwJn3wg12JwYCTat4uHR7sWvEWVnmG_jiEDgNogdg1LQn_Un-qnVTutLxWag$ and include the link in your posting. ***** Thanks for the suggestion Tilo. How long do the viscotears last before evaporating? We are hoping for multiple position imaging over 48 hours. Best, Pablo -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Tilo Moede Sent: Thursday, April 15, 2021 6:15 AM To: [hidden email] Subject: Re: immersion fluid options for long timelapses with long working distance water objective ***** To join, leave or search the confocal microscopy listserv, go to: https://urldefense.com/v3/__http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy__;!!MXfaZl3l!NV0LInRfEiVhwJn3wg12JwYCTat4uHR7sWvEWVnmG_jiEDgNogdg1LQn_Un-qnVTiX8EklI$ Post images on https://urldefense.com/v3/__http://www.imgur.com__;!!MXfaZl3l!NV0LInRfEiVhwJn3wg12JwYCTat4uHR7sWvEWVnmG_jiEDgNogdg1LQn_Un-qnVTutLxWag$ and include the link in your posting. ***** We use viscotears(artificial tear-fluid replacement) as immersion fluid with exactly that lens (and it's 10x and 20x relatives) for in vivo imaging of organoids transplanted to the anterior chamber of the eye with great results.. Something like that should be available from a pharmacy. Cheerio Tilo Hämta Outlook för Android<https://urldefense.com/v3/__https://aka.ms/AAb9ysg__;!!MXfaZl3l!NV0LInRfEiVhwJn3wg12JwYCTat4uHR7sWvEWVnmG_jiEDgNogdg1LQn_Un-qnVTGHe3E_0$ > ________________________________ From: Confocal Microscopy List <[hidden email]> on behalf of Ariel, Pablo <[hidden email]> Sent: Thursday, April 15, 2021 12:05:25 PM To: [hidden email] <[hidden email]> Subject: immersion fluid options for long timelapses with long working distance water objective ***** To join, leave or search the confocal microscopy listserv, go to: https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7CTilo.Moede%40KI.SE%7C3517a286bb174a78639b08d8fff65b64%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637540780863329546%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=2qHml3iWWWFliCmDgStyuaYKjK7jnpvZhSg6vzZxSs8%3D&reserved=0 Post images on https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7CTilo.Moede%40KI.SE%7C3517a286bb174a78639b08d8fff65b64%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637540780863329546%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=rFgQ0fCjIpE%2FNn0EBPQErmRAEpYW6vQ%2BqH37KgH%2FUT0%3D&reserved=0 and include the link in your posting. ***** Dear all, I am demoing a long working distance water objective from Leica (HC FLUOTAR L 25x/0,95 W VISIR, WD: 2.4mm #506375) in an inverted configuration, and am running into some challenges. While the objective is very nice, the long working distance presents a problem for long-duration timelapses with multiple positions. Water works fine, but it eventually evaporates (in hours). Zeiss Immersol W does not evaporate, but cannot hold the column when the objective is over a mm from the coverslip (it drips/slides off). Leica has an automated water dispenser, but it is not available for my current demo. I was wondering if anyone had any ideas for other immersion options that have a surface tension and RI similar to water, but don't evaporate as easily. Thanks, Pablo Pablo Ariel, Ph.D. Assistant Professor Director of the Microscopy Services Laboratory Brinkhous-Bullitt Bldg B04 Department of Pathology and Laboratory Medicine University of North Carolina at Chapel Hill https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.med.unc.edu%2Fmicroscopy&data=04%7C01%7CTilo.Moede%40KI.SE%7C3517a286bb174a78639b08d8fff65b64%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637540780863329546%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=4wo53%2Bh3lqdhhPlnEhu2oibKqKHBI3E2kO%2BBpX4wmfo%3D&reserved=0 Tel: 919-966-2413 När du skickar e-post till Karolinska Institutet (KI) innebär detta att KI kommer att behandla dina personuppgifter. Här finns information om hur KI behandlar personuppgifter<https://urldefense.com/v3/__https://ki.se/medarbetare/integritetsskyddspolicy__;!!MXfaZl3l!NV0LInRfEiVhwJn3wg12JwYCTat4uHR7sWvEWVnmG_jiEDgNogdg1LQn_Un-qnVTMJLmYaA$ >. Sending email to Karolinska Institutet (KI) will result in KI processing your personal data. You can read more about KI's processing of personal data here<https://urldefense.com/v3/__https://ki.se/en/staff/data-protection-policy__;!!MXfaZl3l!NV0LInRfEiVhwJn3wg12JwYCTat4uHR7sWvEWVnmG_jiEDgNogdg1LQn_Un-qnVTobazN_U$ >. |
Tim Feinstein |
In reply to this post by Ariel, Pablo
*****
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. ***** Why not use the Cargille oils that match the refractive index of water? You can get separate oils for matching water at room temperature (~27C) or incubation temperature (37C). I used those for long term imaging without any evaporation issues. I don't remember the product #s but I can look them up if you like. All the best, T Timothy Feinstein, Ph.D. -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Ariel, Pablo Sent: Thursday, April 15, 2021 6:05 AM To: [hidden email] Subject: immersion fluid options for long timelapses with long working distance water objective ***** To join, leave or search the confocal microscopy listserv, go to: https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Ctnf8%40PITT.EDU%7Cb38ac40b056d4715f22c08d8fff65d75%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637540780890462531%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=hA8vGP0sPHELDD2uLakMIMwyzaRT%2FB%2FIzr8W%2BUZTwjQ%3D&reserved=0 Post images on https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Ctnf8%40PITT.EDU%7Cb38ac40b056d4715f22c08d8fff65d75%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637540780890462531%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=ioe%2BTAGtoI5M2ly81aUixXGRZSSMIMp%2FAUipc0RESoM%3D&reserved=0 and include the link in your posting. ***** Dear all, I am demoing a long working distance water objective from Leica (HC FLUOTAR L 25x/0,95 W VISIR, WD: 2.4mm #506375) in an inverted configuration, and am running into some challenges. While the objective is very nice, the long working distance presents a problem for long-duration timelapses with multiple positions. Water works fine, but it eventually evaporates (in hours). Zeiss Immersol W does not evaporate, but cannot hold the column when the objective is over a mm from the coverslip (it drips/slides off). Leica has an automated water dispenser, but it is not available for my current demo. I was wondering if anyone had any ideas for other immersion options that have a surface tension and RI similar to water, but don't evaporate as easily. Thanks, Pablo Pablo Ariel, Ph.D. Assistant Professor Director of the Microscopy Services Laboratory Brinkhous-Bullitt Bldg B04 Department of Pathology and Laboratory Medicine University of North Carolina at Chapel Hill https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.med.unc.edu%2Fmicroscopy&data=04%7C01%7Ctnf8%40PITT.EDU%7Cb38ac40b056d4715f22c08d8fff65d75%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637540780890462531%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=wnOHApiZUAd6ZVqaqBQCrP7O8wCev%2BND97lhM2FmhQY%3D&reserved=0 Tel: 919-966-2413 |
Cammer, Michael-2 |
*****
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. ***** Long working distances require that the fluid maintain its structure, such as with high viscosity or strong surface tension. The oils run down the side of the lens. Cheers- Michael -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Feinstein, Timothy N Sent: Thursday, April 15, 2021 9:55 AM To: [hidden email] Subject: Re: immersion fluid options for long timelapses with long working distance water objective [EXTERNAL] ***** 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. ***** Why not use the Cargille oils that match the refractive index of water? You can get separate oils for matching water at room temperature (~27C) or incubation temperature (37C). I used those for long term imaging without any evaporation issues. I don't remember the product #s but I can look them up if you like. All the best, T Timothy Feinstein, Ph.D. -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Ariel, Pablo Sent: Thursday, April 15, 2021 6:05 AM To: [hidden email] Subject: immersion fluid options for long timelapses with long working distance water objective ***** To join, leave or search the confocal microscopy listserv, go to: https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Ctnf8%40PITT.EDU%7Cb38ac40b056d4715f22c08d8fff65d75%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637540780890462531%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=hA8vGP0sPHELDD2uLakMIMwyzaRT%2FB%2FIzr8W%2BUZTwjQ%3D&reserved=0 Post images on https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Ctnf8%40PITT.EDU%7Cb38ac40b056d4715f22c08d8fff65d75%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637540780890462531%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=ioe%2BTAGtoI5M2ly81aUixXGRZSSMIMp%2FAUipc0RESoM%3D&reserved=0 and include the link in your posting. ***** Dear all, I am demoing a long working distance water objective from Leica (HC FLUOTAR L 25x/0,95 W VISIR, WD: 2.4mm #506375) in an inverted configuration, and am running into some challenges. While the objective is very nice, the long working distance presents a problem for long-duration timelapses with multiple positions. Water works fine, but it eventually evaporates (in hours). Zeiss Immersol W does not evaporate, but cannot hold the column when the objective is over a mm from the coverslip (it drips/slides off). Leica has an automated water dispenser, but it is not available for my current demo. I was wondering if anyone had any ideas for other immersion options that have a surface tension and RI similar to water, but don't evaporate as easily. Thanks, Pablo Pablo Ariel, Ph.D. Assistant Professor Director of the Microscopy Services Laboratory Brinkhous-Bullitt Bldg B04 Department of Pathology and Laboratory Medicine University of North Carolina at Chapel Hill https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.med.unc.edu%2Fmicroscopy&data=04%7C01%7Ctnf8%40PITT.EDU%7Cb38ac40b056d4715f22c08d8fff65d75%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637540780890462531%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=wnOHApiZUAd6ZVqaqBQCrP7O8wCev%2BND97lhM2FmhQY%3D&reserved=0 Tel: 919-966-2413 ------------------------------------------------------------ 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. ================================= |
Periasamy, Ammasi (ap3t)-2 |
In reply to this post by Ariel, Pablo
*****
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. ***** Pablo, Zeiss sells oil with refractive index 1.33. This will go with any companies water immersion lenses. we are using for all the microscopy systems for long time time-lapse imaging. Hope thus helps. Best, Ammasi Dr. Ammasi Periasamy Director and founder of WM Keck Center for Cellular Imaging, Prof. of Biology & Biomed. Eng., University of Virginia, Physical & Life Sciences Building (PLSB 005) 90 Geldard Dr., Charlottesville, VA 22904, USA. http://www.kcci.virginia.edu/people/profile/ap3t Phone: (434) 243-7602 or 982-4869 Fax: (434) 982-5210 E-mail: [hidden email] 20th Annual FLIM and FRET Workshop-March 7-11, 2022. http://www.kcci.virginia.edu/workshop -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Ariel, Pablo Sent: Thursday, April 15, 2021 6:05 AM To: [hidden email] Subject: immersion fluid options for long timelapses with long working distance water objective ***** 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 demoing a long working distance water objective from Leica (HC FLUOTAR L 25x/0,95 W VISIR, WD: 2.4mm #506375) in an inverted configuration, and am running into some challenges. While the objective is very nice, the long working distance presents a problem for long-duration timelapses with multiple positions. Water works fine, but it eventually evaporates (in hours). Zeiss Immersol W does not evaporate, but cannot hold the column when the objective is over a mm from the coverslip (it drips/slides off). Leica has an automated water dispenser, but it is not available for my current demo. I was wondering if anyone had any ideas for other immersion options that have a surface tension and RI similar to water, but don't evaporate as easily. Thanks, Pablo Pablo Ariel, Ph.D. Assistant Professor Director of the Microscopy Services Laboratory Brinkhous-Bullitt Bldg B04 Department of Pathology and Laboratory Medicine University of North Carolina at Chapel Hill http://www.med.unc.edu/microscopy Tel: 919-966-2413 |
Cammer, Michael-2 |
In reply to this post by Tim Feinstein
*****
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. ***** Long working distances require that the fluid maintain its structure, such as with high viscosity or strong surface tension. The oils (at least the ones we've tried such as Immersol W) run down the side of the lens. Cheers- Michael -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Feinstein, Timothy N Sent: Thursday, April 15, 2021 9:55 AM To: [hidden email] Subject: Re: immersion fluid options for long timelapses with long working distance water objective [EXTERNAL] ***** 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. ***** Why not use the Cargille oils that match the refractive index of water? You can get separate oils for matching water at room temperature (~27C) or incubation temperature (37C). I used those for long term imaging without any evaporation issues. I don't remember the product #s but I can look them up if you like. All the best, T Timothy Feinstein, Ph.D. -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Ariel, Pablo Sent: Thursday, April 15, 2021 6:05 AM To: [hidden email] Subject: immersion fluid options for long timelapses with long working distance water objective ***** To join, leave or search the confocal microscopy listserv, go to: https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Ctnf8%40PITT.EDU%7Cb38ac40b056d4715f22c08d8fff65d75%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637540780890462531%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=hA8vGP0sPHELDD2uLakMIMwyzaRT%2FB%2FIzr8W%2BUZTwjQ%3D&reserved=0 Post images on https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Ctnf8%40PITT.EDU%7Cb38ac40b056d4715f22c08d8fff65d75%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637540780890462531%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=ioe%2BTAGtoI5M2ly81aUixXGRZSSMIMp%2FAUipc0RESoM%3D&reserved=0 and include the link in your posting. ***** Dear all, I am demoing a long working distance water objective from Leica (HC FLUOTAR L 25x/0,95 W VISIR, WD: 2.4mm #506375) in an inverted configuration, and am running into some challenges. While the objective is very nice, the long working distance presents a problem for long-duration timelapses with multiple positions. Water works fine, but it eventually evaporates (in hours). Zeiss Immersol W does not evaporate, but cannot hold the column when the objective is over a mm from the coverslip (it drips/slides off). Leica has an automated water dispenser, but it is not available for my current demo. I was wondering if anyone had any ideas for other immersion options that have a surface tension and RI similar to water, but don't evaporate as easily. Thanks, Pablo Pablo Ariel, Ph.D. Assistant Professor Director of the Microscopy Services Laboratory Brinkhous-Bullitt Bldg B04 Department of Pathology and Laboratory Medicine University of North Carolina at Chapel Hill https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.med.unc.edu%2Fmicroscopy&data=04%7C01%7Ctnf8%40PITT.EDU%7Cb38ac40b056d4715f22c08d8fff65d75%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637540780890462531%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=wnOHApiZUAd6ZVqaqBQCrP7O8wCev%2BND97lhM2FmhQY%3D&reserved=0 Tel: 919-966-2413 ------------------------------------------------------------ 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. ================================= |
Ariel, Pablo |
*****
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. ***** Thanks all for the many suggestions received on and off list. I will try the eyedrop approach and let everyone know if we come up with some sort of containment that can reduce evaporation further. Best, Pablo -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Swayne, Theresa C. Sent: Thursday, April 15, 2021 10:10 AM To: [hidden email] Subject: Re: [EXTERNAL] Re: immersion fluid options for long timelapses with long working distance water objective ***** 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 Pablo, We have used Genteal Severe Gel (Hypromellose 0.3%) with our LWD lens on an inverted system. It lasts overnight, but will not last 48 hours without replenishment. For longer experiments, we re-apply during a break in imaging, roughly every 12 hours or so. We also made a little wall out of a glove finger to hold the gel in place and I think this reduces evaporation as well (but it limits ability to do very large Z stacks, as the glove material folds under pressure). Hope this helps, Theresa ------------------------------------ Theresa Swayne, Ph.D. Associate Research Scientist Manager, Confocal and Specialized Microscopy Shared Resource<http://hiccc.columbia.edu/research/sharedresources/confocal> (she/her/hers) Mobile: 347-920-6865 (anytime) [hidden email] <mailto:[hidden email]> (Office hours only: 8-7, Mon-Fri) Core customers: Please contact BOTH Laura Munteanu (elm2157/347-721-7851) AND me for the fastest response. Herbert Irving Comprehensive Cancer Center Columbia University Irving Medical Center Lasker Biomedical Research Building 3960 Broadway, Room 320 New York, NY 10032 From: Confocal Microscopy List <[hidden email]> on behalf of Tilo Moede <[hidden email]> Date: Thursday, April 15, 2021 at 6:35 AM To: [hidden email] <[hidden email]> Subject: [EXTERNAL] Re: immersion fluid options for long timelapses with long working distance water objective ***** 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=DwIFAw&c=G2MiLlal7SXE3PeSnG8W6_JBU6FcdVjSsBSbw6gcR0U&r=JzqbFzGMeaMAlwu5KvYtC8ig-FF8xbvhI6BlFtbXky8&m=4gcujy7AjOTziHxbnfuOrypVxO9f7GuDJ08xTw8y8SY&s=ZZHOPNNkyr5Ym7HuL68QB_BZnfAYLM-vEAmpLIxDOzc&e= Post images on https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwIFAw&c=G2MiLlal7SXE3PeSnG8W6_JBU6FcdVjSsBSbw6gcR0U&r=JzqbFzGMeaMAlwu5KvYtC8ig-FF8xbvhI6BlFtbXky8&m=4gcujy7AjOTziHxbnfuOrypVxO9f7GuDJ08xTw8y8SY&s=PYfF1poS60mJEIWYsRKSmq6N8Q9R1ARdNRUtS-Lc3WY&e= and include the link in your posting. ***** We use viscotears(artificial tear-fluid replacement) as immersion fluid with exactly that lens (and it's 10x and 20x relatives) for in vivo imaging of organoids transplanted to the anterior chamber of the eye with great results.. Something like that should be available from a pharmacy. Cheerio Tilo Hämta Outlook för Android<https://urldefense.proofpoint.com/v2/url?u=https-3A__aka.ms_AAb9ysg&d=DwIFAw&c=G2MiLlal7SXE3PeSnG8W6_JBU6FcdVjSsBSbw6gcR0U&r=JzqbFzGMeaMAlwu5KvYtC8ig-FF8xbvhI6BlFtbXky8&m=4gcujy7AjOTziHxbnfuOrypVxO9f7GuDJ08xTw8y8SY&s=i0M89wHY3rpsoGsdbotuCX1Pus6LNE9KqVCIPdQsbUk&e= > ________________________________ From: Confocal Microscopy List <[hidden email]> on behalf of Ariel, Pablo <[hidden email]> Sent: Thursday, April 15, 2021 12:05:25 PM To: [hidden email] <[hidden email]> Subject: immersion fluid options for long timelapses with long working distance water objective ***** To join, leave or search the confocal microscopy listserv, go to: https://urldefense.proofpoint.com/v2/url?u=https-3A__eur01.safelinks.protection.outlook.com_-3Furl-3Dhttp-253A-252F-252Flists.umn.edu-252Fcgi-2Dbin-252Fwa-253FA0-253Dconfocalmicroscopy-26amp-3Bdata-3D04-257C01-257CTilo.Moede-2540KI.SE-257C3517a286bb174a78639b08d8fff65b64-257Cbff7eef1cf4b4f32be3da1dda043c05d-257C0-257C0-257C637540780863329546-257CUnknown-257CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0-253D-257C1000-26amp-3Bsdata-3D2qHml3iWWWFliCmDgStyuaYKjK7jnpvZhSg6vzZxSs8-253D-26amp-3Breserved-3D0&d=DwIFAw&c=G2MiLlal7SXE3PeSnG8W6_JBU6FcdVjSsBSbw6gcR0U&r=JzqbFzGMeaMAlwu5KvYtC8ig-FF8xbvhI6BlFtbXky8&m=4gcujy7AjOTziHxbnfuOrypVxO9f7GuDJ08xTw8y8SY&s=onGHE-h_33cMxFnQ5PQj8Lk-1aQH-xDHcMIeOYO8Lvw&e= Post images on https://urldefense.proofpoint.com/v2/url?u=https-3A__eur01.safelinks.protection.outlook.com_-3Furl-3Dhttp-253A-252F-252Fwww.imgur.com-252F-26amp-3Bdata-3D04-257C01-257CTilo.Moede-2540KI.SE-257C3517a286bb174a78639b08d8fff65b64-257Cbff7eef1cf4b4f32be3da1dda043c05d-257C0-257C0-257C637540780863329546-257CUnknown-257CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0-253D-257C1000-26amp-3Bsdata-3DrFgQ0fCjIpE-252FNn0EBPQErmRAEpYW6vQ-252BqH37KgH-252FUT0-253D-26amp-3Breserved-3D0&d=DwIFAw&c=G2MiLlal7SXE3PeSnG8W6_JBU6FcdVjSsBSbw6gcR0U&r=JzqbFzGMeaMAlwu5KvYtC8ig-FF8xbvhI6BlFtbXky8&m=4gcujy7AjOTziHxbnfuOrypVxO9f7GuDJ08xTw8y8SY&s=4JAb5EswatQ-lYpnFxzP9qk5OLUH4haCyChomrHYWeM&e= and include the link in your posting. ***** Dear all, I am demoing a long working distance water objective from Leica (HC FLUOTAR L 25x/0,95 W VISIR, WD: 2.4mm #506375) in an inverted configuration, and am running into some challenges. While the objective is very nice, the long working distance presents a problem for long-duration timelapses with multiple positions. Water works fine, but it eventually evaporates (in hours). Zeiss Immersol W does not evaporate, but cannot hold the column when the objective is over a mm from the coverslip (it drips/slides off). Leica has an automated water dispenser, but it is not available for my current demo. I was wondering if anyone had any ideas for other immersion options that have a surface tension and RI similar to water, but don't evaporate as easily. Thanks, Pablo Pablo Ariel, Ph.D. Assistant Professor Director of the Microscopy Services Laboratory Brinkhous-Bullitt Bldg B04 Department of Pathology and Laboratory Medicine University of North Carolina at Chapel Hill https://urldefense.proofpoint.com/v2/url?u=https-3A__eur01.safelinks.protection.outlook.com_-3Furl-3Dhttp-253A-252F-252Fwww.med.unc.edu-252Fmicroscopy-26amp-3Bdata-3D04-257C01-257CTilo.Moede-2540KI.SE-257C3517a286bb174a78639b08d8fff65b64-257Cbff7eef1cf4b4f32be3da1dda043c05d-257C0-257C0-257C637540780863329546-257CUnknown-257CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0-253D-257C1000-26amp-3Bsdata-3D4wo53-252Bh3lqdhhPlnEhu2oibKqKHBI3E2kO-252BBpX4wmfo-253D-26amp-3Breserved-3D0&d=DwIFAw&c=G2MiLlal7SXE3PeSnG8W6_JBU6FcdVjSsBSbw6gcR0U&r=JzqbFzGMeaMAlwu5KvYtC8ig-FF8xbvhI6BlFtbXky8&m=4gcujy7AjOTziHxbnfuOrypVxO9f7GuDJ08xTw8y8SY&s=sPnYkSPIO9WVpX90LDSPI5ylwKf1R1ChYz7qGvkTZ64&e= Tel: 919-966-2413 När du skickar e-post till Karolinska Institutet (KI) innebär detta att KI kommer att behandla dina personuppgifter. Här finns information om hur KI behandlar personuppgifter<https://urldefense.proofpoint.com/v2/url?u=https-3A__ki.se_medarbetare_integritetsskyddspolicy&d=DwIFAw&c=G2MiLlal7SXE3PeSnG8W6_JBU6FcdVjSsBSbw6gcR0U&r=JzqbFzGMeaMAlwu5KvYtC8ig-FF8xbvhI6BlFtbXky8&m=4gcujy7AjOTziHxbnfuOrypVxO9f7GuDJ08xTw8y8SY&s=jrIlbDxhTaT0hy1zcEA35R8Aj6E9B1Ndp-dscg50iPs&e= >. Sending email to Karolinska Institutet (KI) will result in KI processing your personal data. You can read more about KI's processing of personal data here<https://urldefense.proofpoint.com/v2/url?u=https-3A__ki.se_en_staff_data-2Dprotection-2Dpolicy&d=DwIFAw&c=G2MiLlal7SXE3PeSnG8W6_JBU6FcdVjSsBSbw6gcR0U&r=JzqbFzGMeaMAlwu5KvYtC8ig-FF8xbvhI6BlFtbXky8&m=4gcujy7AjOTziHxbnfuOrypVxO9f7GuDJ08xTw8y8SY&s=wIhUo_BGeQ7zzsbSQ_vl-IewAwcVEbKOQmtlzL-7LC0&e= >. |
Cromey, Douglas W - (dcromey)-2 |
*****
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. ***** http://confocal-microscopy-list.588098.n2.nabble.com/water-refill-device-for-WI-objective-td5566734.html I remember that Steve Cody worked on this problem (I met Steve at Jim Pawley's course back in 2008). See his response in the thread above. Doug ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Douglas W. Cromey, M.S. - Associate Scientific Investigator Dept. of Cellular & Molecular Medicine, University of Arizona Life Sciences North, Room 463 1333 N. Martin Ave, Tucson, AZ 85721 USA office: LSN 463 email: [hidden email] voice: 520-626-2824 fax: 520-626-2097 UA Microscopy Alliance - http://microscopy.arizona.edu/ A collaborative effort to bring information about shared microscopy facilities to the University of Arizona and the community. -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Ariel, Pablo Sent: Thursday, April 15, 2021 7:49 AM To: [hidden email] Subject: [EXT]Re: [EXTERNAL] Re: immersion fluid options for long timelapses with long working distance water objective External Email ***** 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. ***** Thanks all for the many suggestions received on and off list. I will try the eyedrop approach and let everyone know if we come up with some sort of containment that can reduce evaporation further. Best, Pablo -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Swayne, Theresa C. Sent: Thursday, April 15, 2021 10:10 AM To: [hidden email] Subject: Re: [EXTERNAL] Re: immersion fluid options for long timelapses with long working distance water objective ***** 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 Pablo, We have used Genteal Severe Gel (Hypromellose 0.3%) with our LWD lens on an inverted system. It lasts overnight, but will not last 48 hours without replenishment. For longer experiments, we re-apply during a break in imaging, roughly every 12 hours or so. We also made a little wall out of a glove finger to hold the gel in place and I think this reduces evaporation as well (but it limits ability to do very large Z stacks, as the glove material folds under pressure). Hope this helps, Theresa ------------------------------------ Theresa Swayne, Ph.D. Associate Research Scientist Manager, Confocal and Specialized Microscopy Shared Resource<http://hiccc.columbia.edu/research/sharedresources/confocal> (she/her/hers) Mobile: 347-920-6865 (anytime) [hidden email] <mailto:[hidden email]> (Office hours only: 8-7, Mon-Fri) Core customers: Please contact BOTH Laura Munteanu (elm2157/347-721-7851) AND me for the fastest response. Herbert Irving Comprehensive Cancer Center Columbia University Irving Medical Center Lasker Biomedical Research Building 3960 Broadway, Room 320 New York, NY 10032 From: Confocal Microscopy List <[hidden email]> on behalf of Tilo Moede <[hidden email]> Date: Thursday, April 15, 2021 at 6:35 AM To: [hidden email] <[hidden email]> Subject: [EXTERNAL] Re: immersion fluid options for long timelapses with long working distance water objective ***** 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=DwIFAw&c=G2MiLlal7SXE3PeSnG8W6_JBU6FcdVjSsBSbw6gcR0U&r=JzqbFzGMeaMAlwu5KvYtC8ig-FF8xbvhI6BlFtbXky8&m=4gcujy7AjOTziHxbnfuOrypVxO9f7GuDJ08xTw8y8SY&s=ZZHOPNNkyr5Ym7HuL68QB_BZnfAYLM-vEAmpLIxDOzc&e= Post images on https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwIFAw&c=G2MiLlal7SXE3PeSnG8W6_JBU6FcdVjSsBSbw6gcR0U&r=JzqbFzGMeaMAlwu5KvYtC8ig-FF8xbvhI6BlFtbXky8&m=4gcujy7AjOTziHxbnfuOrypVxO9f7GuDJ08xTw8y8SY&s=PYfF1poS60mJEIWYsRKSmq6N8Q9R1ARdNRUtS-Lc3WY&e= and include the link in your posting. ***** We use viscotears(artificial tear-fluid replacement) as immersion fluid with exactly that lens (and it's 10x and 20x relatives) for in vivo imaging of organoids transplanted to the anterior chamber of the eye with great results.. Something like that should be available from a pharmacy. Cheerio Tilo Hämta Outlook för Android<https://urldefense.proofpoint.com/v2/url?u=https-3A__aka.ms_AAb9ysg&d=DwIFAw&c=G2MiLlal7SXE3PeSnG8W6_JBU6FcdVjSsBSbw6gcR0U&r=JzqbFzGMeaMAlwu5KvYtC8ig-FF8xbvhI6BlFtbXky8&m=4gcujy7AjOTziHxbnfuOrypVxO9f7GuDJ08xTw8y8SY&s=i0M89wHY3rpsoGsdbotuCX1Pus6LNE9KqVCIPdQsbUk&e= > ________________________________ From: Confocal Microscopy List <[hidden email]> on behalf of Ariel, Pablo <[hidden email]> Sent: Thursday, April 15, 2021 12:05:25 PM To: [hidden email] <[hidden email]> Subject: immersion fluid options for long timelapses with long working distance water objective ***** To join, leave or search the confocal microscopy listserv, go to: https://urldefense.proofpoint.com/v2/url?u=https-3A__eur01.safelinks.protection.outlook.com_-3Furl-3Dhttp-253A-252F-252Flists.umn.edu-252Fcgi-2Dbin-252Fwa-253FA0-253Dconfocalmicroscopy-26amp-3Bdata-3D04-257C01-257CTilo.Moede-2540KI.SE-257C3517a286bb174a78639b08d8fff65b64-257Cbff7eef1cf4b4f32be3da1dda043c05d-257C0-257C0-257C637540780863329546-257CUnknown-257CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0-253D-257C1000-26amp-3Bsdata-3D2qHml3iWWWFliCmDgStyuaYKjK7jnpvZhSg6vzZxSs8-253D-26amp-3Breserved-3D0&d=DwIFAw&c=G2MiLlal7SXE3PeSnG8W6_JBU6FcdVjSsBSbw6gcR0U&r=JzqbFzGMeaMAlwu5KvYtC8ig-FF8xbvhI6BlFtbXky8&m=4gcujy7AjOTziHxbnfuOrypVxO9f7GuDJ08xTw8y8SY&s=onGHE-h_33cMxFnQ5PQj8Lk-1aQH-xDHcMIeOYO8Lvw&e= Post images on https://urldefense.proofpoint.com/v2/url?u=https-3A__eur01.safelinks.protection.outlook.com_-3Furl-3Dhttp-253A-252F-252Fwww.imgur.com-252F-26amp-3Bdata-3D04-257C01-257CTilo.Moede-2540KI.SE-257C3517a286bb174a78639b08d8fff65b64-257Cbff7eef1cf4b4f32be3da1dda043c05d-257C0-257C0-257C637540780863329546-257CUnknown-257CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0-253D-257C1000-26amp-3Bsdata-3DrFgQ0fCjIpE-252FNn0EBPQErmRAEpYW6vQ-252BqH37KgH-252FUT0-253D-26amp-3Breserved-3D0&d=DwIFAw&c=G2MiLlal7SXE3PeSnG8W6_JBU6FcdVjSsBSbw6gcR0U&r=JzqbFzGMeaMAlwu5KvYtC8ig-FF8xbvhI6BlFtbXky8&m=4gcujy7AjOTziHxbnfuOrypVxO9f7GuDJ08xTw8y8SY&s=4JAb5EswatQ-lYpnFxzP9qk5OLUH4haCyChomrHYWeM&e= and include the link in your posting. ***** Dear all, I am demoing a long working distance water objective from Leica (HC FLUOTAR L 25x/0,95 W VISIR, WD: 2.4mm #506375) in an inverted configuration, and am running into some challenges. While the objective is very nice, the long working distance presents a problem for long-duration timelapses with multiple positions. Water works fine, but it eventually evaporates (in hours). Zeiss Immersol W does not evaporate, but cannot hold the column when the objective is over a mm from the coverslip (it drips/slides off). Leica has an automated water dispenser, but it is not available for my current demo. I was wondering if anyone had any ideas for other immersion options that have a surface tension and RI similar to water, but don't evaporate as easily. Thanks, Pablo Pablo Ariel, Ph.D. Assistant Professor Director of the Microscopy Services Laboratory Brinkhous-Bullitt Bldg B04 Department of Pathology and Laboratory Medicine University of North Carolina at Chapel Hill https://urldefense.proofpoint.com/v2/url?u=https-3A__eur01.safelinks.protection.outlook.com_-3Furl-3Dhttp-253A-252F-252Fwww.med.unc.edu-252Fmicroscopy-26amp-3Bdata-3D04-257C01-257CTilo.Moede-2540KI.SE-257C3517a286bb174a78639b08d8fff65b64-257Cbff7eef1cf4b4f32be3da1dda043c05d-257C0-257C0-257C637540780863329546-257CUnknown-257CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0-253D-257C1000-26amp-3Bsdata-3D4wo53-252Bh3lqdhhPlnEhu2oibKqKHBI3E2kO-252BBpX4wmfo-253D-26amp-3Breserved-3D0&d=DwIFAw&c=G2MiLlal7SXE3PeSnG8W6_JBU6FcdVjSsBSbw6gcR0U&r=JzqbFzGMeaMAlwu5KvYtC8ig-FF8xbvhI6BlFtbXky8&m=4gcujy7AjOTziHxbnfuOrypVxO9f7GuDJ08xTw8y8SY&s=sPnYkSPIO9WVpX90LDSPI5ylwKf1R1ChYz7qGvkTZ64&e= Tel: 919-966-2413 När du skickar e-post till Karolinska Institutet (KI) innebär detta att KI kommer att behandla dina personuppgifter. Här finns information om hur KI behandlar personuppgifter<https://urldefense.proofpoint.com/v2/url?u=https-3A__ki.se_medarbetare_integritetsskyddspolicy&d=DwIFAw&c=G2MiLlal7SXE3PeSnG8W6_JBU6FcdVjSsBSbw6gcR0U&r=JzqbFzGMeaMAlwu5KvYtC8ig-FF8xbvhI6BlFtbXky8&m=4gcujy7AjOTziHxbnfuOrypVxO9f7GuDJ08xTw8y8SY&s=jrIlbDxhTaT0hy1zcEA35R8Aj6E9B1Ndp-dscg50iPs&e= >. Sending email to Karolinska Institutet (KI) will result in KI processing your personal data. You can read more about KI's processing of personal data here<https://urldefense.proofpoint.com/v2/url?u=https-3A__ki.se_en_staff_data-2Dprotection-2Dpolicy&d=DwIFAw&c=G2MiLlal7SXE3PeSnG8W6_JBU6FcdVjSsBSbw6gcR0U&r=JzqbFzGMeaMAlwu5KvYtC8ig-FF8xbvhI6BlFtbXky8&m=4gcujy7AjOTziHxbnfuOrypVxO9f7GuDJ08xTw8y8SY&s=wIhUo_BGeQ7zzsbSQ_vl-IewAwcVEbKOQmtlzL-7LC0&e= >. |
Jeremy Adler-4 |
In reply to this post by Tim Feinstein
*****
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. ***** Assuming you are using a petri dish with a glass bottom and adherent cells. Grow you cells on a second coverslip and mount it in the petri dish face down with a 1.5mm spacer underneath. This should get the objective close enough to the bottom of the petri dish for the oil to occupy the gap. Jeremy Adler BioVis Uppsala -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Feinstein, Timothy N Sent: Thursday, April 15, 2021 3:55 PM To: [hidden email] Subject: Re: immersion fluid options for long timelapses with long working distance water objective ***** 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. ***** Why not use the Cargille oils that match the refractive index of water? You can get separate oils for matching water at room temperature (~27C) or incubation temperature (37C). I used those for long term imaging without any evaporation issues. I don't remember the product #s but I can look them up if you like. All the best, T Timothy Feinstein, Ph.D. -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Ariel, Pablo Sent: Thursday, April 15, 2021 6:05 AM To: [hidden email] Subject: immersion fluid options for long timelapses with long working distance water objective ***** To join, leave or search the confocal microscopy listserv, go to: https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Ctnf8%40PITT.EDU%7Cb38ac40b056d4715f22c08d8fff65d75%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637540780890462531%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=hA8vGP0sPHELDD2uLakMIMwyzaRT%2FB%2FIzr8W%2BUZTwjQ%3D&reserved=0 Post images on https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Ctnf8%40PITT.EDU%7Cb38ac40b056d4715f22c08d8fff65d75%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637540780890462531%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=ioe%2BTAGtoI5M2ly81aUixXGRZSSMIMp%2FAUipc0RESoM%3D&reserved=0 and include the link in your posting. ***** Dear all, I am demoing a long working distance water objective from Leica (HC FLUOTAR L 25x/0,95 W VISIR, WD: 2.4mm #506375) in an inverted configuration, and am running into some challenges. While the objective is very nice, the long working distance presents a problem for long-duration timelapses with multiple positions. Water works fine, but it eventually evaporates (in hours). Zeiss Immersol W does not evaporate, but cannot hold the column when the objective is over a mm from the coverslip (it drips/slides off). Leica has an automated water dispenser, but it is not available for my current demo. I was wondering if anyone had any ideas for other immersion options that have a surface tension and RI similar to water, but don't evaporate as easily. Thanks, Pablo Pablo Ariel, Ph.D. Assistant Professor Director of the Microscopy Services Laboratory Brinkhous-Bullitt Bldg B04 Department of Pathology and Laboratory Medicine University of North Carolina at Chapel Hill https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.med.unc.edu%2Fmicroscopy&data=04%7C01%7Ctnf8%40PITT.EDU%7Cb38ac40b056d4715f22c08d8fff65d75%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637540780890462531%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=wnOHApiZUAd6ZVqaqBQCrP7O8wCev%2BND97lhM2FmhQY%3D&reserved=0 Tel: 919-966-2413 När du har kontakt med oss på Uppsala universitet med e-post så innebär det att vi behandlar dina personuppgifter. För att läsa mer om hur vi gör det kan du läsa här: http://www.uu.se/om-uu/dataskydd-personuppgifter/ E-mailing Uppsala University means that we will process your personal data. For more information on how this is performed, please read here: http://www.uu.se/en/about-uu/data-protection-policy |
Michael Giacomelli-2 |
In reply to this post by Ariel, Pablo
*****
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 use UV cure adhesive to make a wall around the coverslip and then fill up the coverslip with a larger volume of water. It'll eventually evaporate, but for <24 hours it isn't too hard to have enough water that evaporation isn't a problem. It is not an elegant solution but it has always worked well enough. Michael On Thu, Apr 15, 2021 at 6:08 AM Ariel, Pablo <[hidden email]> wrote: > > ***** > 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=DwIFAg&c=kbmfwr1Yojg42sGEpaQh5ofMHBeTl9EI2eaqQZhHbOU&r=0LyF_z8oU1XGGyisIeOIXyIGIM5IYb3NcLjxHjUca5Y&m=b0tDWzqI-CVUzLk7XzsOt3qQqQtXQsPpKR__Jwh_go8&s=mZn0L-XgBz4bsBzZdtkcMvCe8mIRaV2xL8RwpW1SIOE&e= > Post images on https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwIFAg&c=kbmfwr1Yojg42sGEpaQh5ofMHBeTl9EI2eaqQZhHbOU&r=0LyF_z8oU1XGGyisIeOIXyIGIM5IYb3NcLjxHjUca5Y&m=b0tDWzqI-CVUzLk7XzsOt3qQqQtXQsPpKR__Jwh_go8&s=F-0sPOkToi58KLNeOYOsyYK_7bfx-_y3pF3JLIIPpkE&e= and include the link in your posting. > ***** > > Dear all, > > I am demoing a long working distance water objective from Leica (HC FLUOTAR L 25x/0,95 W VISIR, WD: 2.4mm #506375) in an inverted configuration, and am running into some challenges. > > While the objective is very nice, the long working distance presents a problem for long-duration timelapses with multiple positions. Water works fine, but it eventually evaporates (in hours). Zeiss Immersol W does not evaporate, but cannot hold the column when the objective is over a mm from the coverslip (it drips/slides off). Leica has an automated water dispenser, but it is not available for my current demo. I was wondering if anyone had any ideas for other immersion options that have a surface tension and RI similar to water, but don't evaporate as easily. > > Thanks, > Pablo > > Pablo Ariel, Ph.D. > Assistant Professor > Director of the Microscopy Services Laboratory > Brinkhous-Bullitt Bldg B04 > Department of Pathology and Laboratory Medicine > University of North Carolina at Chapel Hill > https://urldefense.proofpoint.com/v2/url?u=http-3A__www.med.unc.edu_microscopy&d=DwIFAg&c=kbmfwr1Yojg42sGEpaQh5ofMHBeTl9EI2eaqQZhHbOU&r=0LyF_z8oU1XGGyisIeOIXyIGIM5IYb3NcLjxHjUca5Y&m=b0tDWzqI-CVUzLk7XzsOt3qQqQtXQsPpKR__Jwh_go8&s=EYNR-YcjTsR3JTlVsAg8U4yVx_EiDEhR7y1rLbo-YsA&e= > Tel: 919-966-2413 |
Craig Brideau |
In reply to this post by Jeremy Adler-4
*****
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. ***** Leica also sells a watering system. One of the cores around here has it on their SP8. It seems to work fairly well; slowly replenishing water as it evaporates and wicking away any excess with a foam collar fitting. Craig On Thu, Apr 15, 2021 at 10:58 AM Jeremy Adler <[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. > ***** > > Assuming you are using a petri dish with a glass bottom and adherent cells. > Grow you cells on a second coverslip and mount it in the petri dish face > down with a 1.5mm spacer underneath. > This should get the objective close enough to the bottom of the petri dish > for the oil to occupy the gap. > > Jeremy Adler > BioVis > Uppsala > > > -----Original Message----- > From: Confocal Microscopy List <[hidden email]> On > Behalf Of Feinstein, Timothy N > Sent: Thursday, April 15, 2021 3:55 PM > To: [hidden email] > Subject: Re: immersion fluid options for long timelapses with long working > distance water objective > > ***** > 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. > ***** > > Why not use the Cargille oils that match the refractive index of water? > You can get separate oils for matching water at room temperature (~27C) or > incubation temperature (37C). I used those for long term imaging without > any evaporation issues. I don't remember the product #s but I can look > them up if you like. > > All the best, > > > T > > Timothy Feinstein, Ph.D. > > > -----Original Message----- > From: Confocal Microscopy List <[hidden email]> On > Behalf Of Ariel, Pablo > Sent: Thursday, April 15, 2021 6:05 AM > To: [hidden email] > Subject: immersion fluid options for long timelapses with long working > distance water objective > > ***** > To join, leave or search the confocal microscopy listserv, go to: > > https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Ctnf8%40PITT.EDU%7Cb38ac40b056d4715f22c08d8fff65d75%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637540780890462531%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=hA8vGP0sPHELDD2uLakMIMwyzaRT%2FB%2FIzr8W%2BUZTwjQ%3D&reserved=0 > Post images on > https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Ctnf8%40PITT.EDU%7Cb38ac40b056d4715f22c08d8fff65d75%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637540780890462531%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=ioe%2BTAGtoI5M2ly81aUixXGRZSSMIMp%2FAUipc0RESoM%3D&reserved=0 > and include the link in your posting. > ***** > > Dear all, > > I am demoing a long working distance water objective from Leica (HC > FLUOTAR L 25x/0,95 W VISIR, WD: 2.4mm #506375) in an inverted > configuration, and am running into some challenges. > > While the objective is very nice, the long working distance presents a > problem for long-duration timelapses with multiple positions. Water works > fine, but it eventually evaporates (in hours). Zeiss Immersol W does not > evaporate, but cannot hold the column when the objective is over a mm from > the coverslip (it drips/slides off). Leica has an automated water > dispenser, but it is not available for my current demo. I was wondering if > anyone had any ideas for other immersion options that have a surface > tension and RI similar to water, but don't evaporate as easily. > > Thanks, > Pablo > > Pablo Ariel, Ph.D. > Assistant Professor > Director of the Microscopy Services Laboratory Brinkhous-Bullitt Bldg B04 > Department of Pathology and Laboratory Medicine University of North > Carolina at Chapel Hill > > https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.med.unc.edu%2Fmicroscopy&data=04%7C01%7Ctnf8%40PITT.EDU%7Cb38ac40b056d4715f22c08d8fff65d75%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637540780890462531%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=wnOHApiZUAd6ZVqaqBQCrP7O8wCev%2BND97lhM2FmhQY%3D&reserved=0 > Tel: 919-966-2413 > > > > > > > > > När du har kontakt med oss på Uppsala universitet med e-post så innebär > det att vi behandlar dina personuppgifter. För att läsa mer om hur vi gör > det kan du läsa här: http://www.uu.se/om-uu/dataskydd-personuppgifter/ > > E-mailing Uppsala University means that we will process your personal > data. For more information on how this is performed, please read here: > http://www.uu.se/en/about-uu/data-protection-policy > |
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