Glycerol/Type G immersion liquid?
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JanTonnesen |
Glycerol/Type G immersion liquid?
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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. ***** Hello everyone Does anyone know if glycerol immersion liquid is in fact merely glycerol, and off-the-shelf (molecular biology grade) glycerol be used instead of designated glycerol immersion liquid? If so, can the refractive index be lowered simply by adding water and thoroughly vortexing/sonicating to get immersion liquid similar to Type G? Asking for a friend, obviously. All the best, and thanks in advance, Jan |
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Nicolai.Urban@mpfi.org |
Re: Glycerol/Type G immersion liquid?
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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 Jan, I have in the past used various different solutions as immersion liquids for my glycerol immersion lens. You can straight up use glycerol, but with a refractive index of n=1.47 you ideally want to bring it down (with water) to n=1.45 to match the specifications of the lens. You can correct for some mismatch using the correction collar, but this takes quite some time and practice, so best keep it to a minimum, if possible. This works perfectly fine, with two main caveats. First, over time the water in the mixture evaporates, causing the refractive index to tick up slowly. This induces increasing aberrations, or requires frequent AO, correction collar readjustment, or reapplication of fresh immersion liquid - all of which you would like to avoid when performing live imaging, especially. If I am correct, branded immersion liquids (e.g. Leica Type G) include additives aimed at slowing the evaporation of water. This would slow the process of RI change, requiring less frequent realignment. Also, this means the refractive index of your "stock" immersion liquid is more constant and predictable, leading to less time spent on pre-aligning your correction collar/AO at the beginning of an experiment. Especially if your vial of pre-mixed immersion liquid sits on a shelf for long periods of time in between imaging... That being said, I have imaged with both, and you can easily make both work. Especially if you want to tweak your RI a little to pre-compensate aberrations (such as are incurred by imaging deeper within living tissue), this is best done by either pre-mixing your immersion liquids yourself, or by simply buying RI-matched liquids with the appropriate RI. I have used Cargille immersion liquids in a range of RIs to to this, and they worked fine. They are slightly pungent, however, and can be more unpleasant to work with. Hope this helps! Feel free to contact me directly if you want anything more in-depth. Best of luck, Nicolai >>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>> Nicolai T. Urban, Ph.D. MPFI Imaging Center – Light Microscopy Core Max Planck Florida Institute for Neuroscience One Max Planck Way, Jupiter FL 33418 -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Jan Tonnesen Sent: Tuesday, 23 June 2020 08:30 To: [hidden email] Subject: Glycerol/Type G immersion liquid? ***** To join, leave or search the confocal microscopy listserv, go to: https://nam03.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=02%7C01%7CNicolai.Urban%40MPFI.ORG%7C69800ad9421e4291139908d81771337a%7C947b45517db44636a5fd1bdcad603ed0%7C0%7C1%7C637285122259605249&sdata=xrkeVAkj7iYBv%2FTaG9u6LowxbwqkQKMfEUzWQNVv8a4%3D&reserved=0 Post images on https://nam03.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=02%7C01%7CNicolai.Urban%40MPFI.ORG%7C69800ad9421e4291139908d81771337a%7C947b45517db44636a5fd1bdcad603ed0%7C0%7C1%7C637285122259605249&sdata=nY3zRhxyGlFoF1OYb40%2FflQQq1xhKvH0SF9whIi%2B26c%3D&reserved=0 and include the link in your posting. ***** Hello everyone Does anyone know if glycerol immersion liquid is in fact merely glycerol, and off-the-shelf (molecular biology grade) glycerol be used instead of designated glycerol immersion liquid? If so, can the refractive index be lowered simply by adding water and thoroughly vortexing/sonicating to get immersion liquid similar to Type G? Asking for a friend, obviously. All the best, and thanks in advance, Jan |
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Konstantín Levitskiy |
Re: Glycerol/Type G immersion liquid?
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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 Jan. We use glycerol objectives for many years. The advice from Leica always was glicerol/water 80:20. Best regards, Konstantin ________________________________ De: Confocal Microscopy List <[hidden email]> en nombre de [hidden email] <[hidden email]> Enviado: martes, 23 de junio de 2020 20:51 Para: [hidden email] <[hidden email]> Asunto: Re: Glycerol/Type G immersion liquid? ***** 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 Jan, I have in the past used various different solutions as immersion liquids for my glycerol immersion lens. You can straight up use glycerol, but with a refractive index of n=1.47 you ideally want to bring it down (with water) to n=1.45 to match the specifications of the lens. You can correct for some mismatch using the correction collar, but this takes quite some time and practice, so best keep it to a minimum, if possible. This works perfectly fine, with two main caveats. First, over time the water in the mixture evaporates, causing the refractive index to tick up slowly. This induces increasing aberrations, or requires frequent AO, correction collar readjustment, or reapplication of fresh immersion liquid - all of which you would like to avoid when performing live imaging, especially. If I am correct, branded immersion liquids (e.g. Leica Type G) include additives aimed at slowing the evaporation of water. This would slow the process of RI change, requiring less frequent realignment. Also, this means the refractive index of your "stock" immersion liquid is more constant and predictable, leading to less time spent on pre-aligning your correction collar/AO at the beginning of an experiment. Especially if your vial of pre-mixed immersion liquid sits on a shelf for long periods of time in between imaging... That being said, I have imaged with both, and you can easily make both work. Especially if you want to tweak your RI a little to pre-compensate aberrations (such as are incurred by imaging deeper within living tissue), this is best done by either pre-mixing your immersion liquids yourself, or by simply buying RI-matched liquids with the appropriate RI. I have used Cargille immersion liquids in a range of RIs to to this, and they worked fine. They are slightly pungent, however, and can be more unpleasant to work with. Hope this helps! Feel free to contact me directly if you want anything more in-depth. Best of luck, Nicolai >>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>> Nicolai T. Urban, Ph.D. MPFI Imaging Center – Light Microscopy Core Max Planck Florida Institute for Neuroscience One Max Planck Way, Jupiter FL 33418 -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Jan Tonnesen Sent: Tuesday, 23 June 2020 08:30 To: [hidden email] Subject: Glycerol/Type G immersion liquid? ***** To join, leave or search the confocal microscopy listserv, go to: https://nam03.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=02%7C01%7CNicolai.Urban%40MPFI.ORG%7C69800ad9421e4291139908d81771337a%7C947b45517db44636a5fd1bdcad603ed0%7C0%7C1%7C637285122259605249&sdata=xrkeVAkj7iYBv%2FTaG9u6LowxbwqkQKMfEUzWQNVv8a4%3D&reserved=0 Post images on https://nam03.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=02%7C01%7CNicolai.Urban%40MPFI.ORG%7C69800ad9421e4291139908d81771337a%7C947b45517db44636a5fd1bdcad603ed0%7C0%7C1%7C637285122259605249&sdata=nY3zRhxyGlFoF1OYb40%2FflQQq1xhKvH0SF9whIi%2B26c%3D&reserved=0 and include the link in your posting. ***** Hello everyone Does anyone know if glycerol immersion liquid is in fact merely glycerol, and off-the-shelf (molecular biology grade) glycerol be used instead of designated glycerol immersion liquid? If so, can the refractive index be lowered simply by adding water and thoroughly vortexing/sonicating to get immersion liquid similar to Type G? Asking for a friend, obviously. All the best, and thanks in advance, Jan |
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