Tissue fixations for safe and functional COVID-19 imaging
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Mark A. Sanders |
Tissue fixations for safe and functional COVID-19 imaging
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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. ***** Greetings Colleagues, I hope this email request finds you well. Many of our shared imaging resources may be requested to image COVID-19 infected tissues samples. Samples containing live virus are at least a BSL-3 situation so many labs must successfully fix the tissue to inactivate the virus to a manageable safety level while preserving the molecular integrity associated with the assay prior to the imaging steps. As is historically the case, there may not be a single fixation method for all assays, but they all must inactivate the virus for safe downstream steps. There are many options out there for viral inactivation methods for biological samples, including this one from WHO concerning the ebola virus ( https://www.paho.org/hq/dmdocuments/2014/2014-cha-procedures-inactivation-ebola.pdf ) Has anyone come across preferred infected tissue fixation methods for COVID-19 samples for either FFPE or fixed cryosectioned tissues? References mandatory. Perhaps many of us will find the answer useful to either safely perform assays or to not do them at all based on our circumstances. Stay healthy, Mark *Working Remotely M-F as needed, (available via email)* -- Mark Sanders University of Minnesota Program Director Twin Cities Campus University Imaging Centers Nikon Center of Excellence <https://www.nikoninstruments.com/Imaging-Centers/Nikon-Centers-of-Excellence/Nikon-CofE-Locations/Americas/University-of-Minnesota> uic.umn.edu |
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Christoph Ruediger Bauer |
Re: Tissue fixations for safe and functional COVID-19 imaging
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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. ***** Dear all, I have been asked to propose a protocol to fix Covid-19 infected cultured cells for fluorescence observations and for TEM embedding. I remembered to have seen Marks question (see below) and downloaded the PDF he is mentioning. Did anybody find and maybe tested in between additional protocols that could be recommended? Thanks a lot and best wishes, Christoph BIOIMAGING CENTER University of Geneva - Science II Room 245 30, Quai Ernest Ansermet CH - 1211 Genève 4 Dr. Christoph R. Bauer -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Mark Sanders Sent: 16 April 2020 19:03 To: [hidden email] Subject: Tissue fixations for safe and functional COVID-19 imaging ***** 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. ***** Greetings Colleagues, I hope this email request finds you well. Many of our shared imaging resources may be requested to image COVID-19 infected tissues samples. Samples containing live virus are at least a BSL-3 situation so many labs must successfully fix the tissue to inactivate the virus to a manageable safety level while preserving the molecular integrity associated with the assay prior to the imaging steps. As is historically the case, there may not be a single fixation method for all assays, but they all must inactivate the virus for safe downstream steps. There are many options out there for viral inactivation methods for biological samples, including this one from WHO concerning the ebola virus ( https://www.paho.org/hq/dmdocuments/2014/2014-cha-procedures-inactivation-ebola.pdf ) Has anyone come across preferred infected tissue fixation methods for COVID-19 samples for either FFPE or fixed cryosectioned tissues? References mandatory. Perhaps many of us will find the answer useful to either safely perform assays or to not do them at all based on our circumstances. Stay healthy, Mark *Working Remotely M-F as needed, (available via email)* -- Mark Sanders University of Minnesota Program Director Twin Cities Campus University Imaging Centers Nikon Center of Excellence <https://www.nikoninstruments.com/Imaging-Centers/Nikon-Centers-of-Excellence/Nikon-CofE-Locations/Americas/University-of-Minnesota> uic.umn.edu |
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Pippa Hawes-2 |
Re: [EXTERNAL] Re: Tissue fixations for safe and functional COVID-19 imaging
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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. ***** Dear all We are running a seminar that touches on these issues this Friday 22 May 13:00 (British Summer Time). Here is the link for registration if anyone is interested: https://www.crick.ac.uk/whats-on/webinarimaging-sars-cov-2-safely-protecting-the-uk-microscopy-community Thanks Pippa Prof Pippa Hawes FRMS Head of Bioimaging t: +44 (0)1483 231026 e: [hidden email] The Pirbright Institute, Ash Road, Pirbright, Woking, Surrey GU24 0NF t: +44 (0)1483 232441 f: +44 (0)1483 232448 e: [hidden email] w: www.pirbright.ac.uk -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Christoph Ruediger Bauer Sent: 19 May 2020 15:15 To: [hidden email] Subject: [EXTERNAL] Re: Tissue fixations for safe and functional COVID-19 imaging ***** 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 have been asked to propose a protocol to fix Covid-19 infected cultured cells for fluorescence observations and for TEM embedding. I remembered to have seen Marks question (see below) and downloaded the PDF he is mentioning. Did anybody find and maybe tested in between additional protocols that could be recommended? Thanks a lot and best wishes, Christoph BIOIMAGING CENTER University of Geneva - Science II Room 245 30, Quai Ernest Ansermet CH - 1211 Genève 4 Dr. Christoph R. Bauer -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Mark Sanders Sent: 16 April 2020 19:03 To: [hidden email] Subject: Tissue fixations for safe and functional COVID-19 imaging ***** 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. ***** Greetings Colleagues, I hope this email request finds you well. Many of our shared imaging resources may be requested to image COVID-19 infected tissues samples. Samples containing live virus are at least a BSL-3 situation so many labs must successfully fix the tissue to inactivate the virus to a manageable safety level while preserving the molecular integrity associated with the assay prior to the imaging steps. As is historically the case, there may not be a single fixation method for all assays, but they all must inactivate the virus for safe downstream steps. There are many options out there for viral inactivation methods for biological samples, including this one from WHO concerning the ebola virus ( https://www.paho.org/hq/dmdocuments/2014/2014-cha-procedures-inactivation-ebola.pdf ) Has anyone come across preferred infected tissue fixation methods for COVID-19 samples for either FFPE or fixed cryosectioned tissues? References mandatory. Perhaps many of us will find the answer useful to either safely perform assays or to not do them at all based on our circumstances. Stay healthy, Mark *Working Remotely M-F as needed, (available via email)* -- Mark Sanders University of Minnesota Program Director Twin Cities Campus University Imaging Centers Nikon Center of Excellence <https://www.nikoninstruments.com/Imaging-Centers/Nikon-Centers-of-Excellence/Nikon-CofE-Locations/Americas/University-of-Minnesota> uic.umn.edu ________________________________ The Pirbright Institute receives strategic funding from BBSRC. The information contained in this message may be confidential or legally privileged and is intended solely for the addressee. If you have received this message in error please delete it & notify the originator immediately. Unauthorised use, disclosure, copying or alteration of this message is forbidden & may be unlawful. The contents of this e-mail are the views of the sender and do not necessarily represent the views of the Institute. This email and associated attachments has been checked locally for viruses but we can accept no responsibility once it has left our systems. Communications on Institute computers are monitored to secure the effective operation of the systems and for other lawful purposes. The Pirbright Institute is a company limited by guarantee, registered in England no. 559784. The Institute is also a registered charity. |
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Carol Heckman |
Re: Tissue fixations for safe and functional COVID-19 imaging
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In reply to this post by Christoph Ruediger Bauer
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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. ***** That's a demanding task. To conserve antigenicity, you cannot fix with glutaraldehyde, at least not much glutaraldehyde, and to get good morphology you almost have to have some glutaraldehyde. Antigens vary in their degree of resistance to glutaraldehyde-induced inactivation. If you could split the sample in two, and process one half for immunofluorescence and the other for TEM, that would work best. Carol Heckman Bowling Green State University ________________________________ From: Confocal Microscopy List <[hidden email]> on behalf of Christoph Ruediger Bauer <[hidden email]> Sent: Tuesday, May 19, 2020 10:15 AM To: [hidden email] <[hidden email]> Subject: [EXTERNAL] Re: Tissue fixations for safe and functional COVID-19 imaging ***** To join, leave or search the confocal microscopy listserv, go to: https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=02%7C01%7Checkman%40bgsu.edu%7C0c57ad04507c4a1c4ce308d7fbff3fda%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254946012393579&sdata=AUOl7fqO26QSreknKotLJptmaEUrsk70wppcSNTsICs%3D&reserved=0 Post images on https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=02%7C01%7Checkman%40bgsu.edu%7C0c57ad04507c4a1c4ce308d7fbff3fda%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254946012393579&sdata=8S7c8RtZ8axAyZQvoCrVanS01Qk%2BmUQwxj8uaxwhsAk%3D&reserved=0 and include the link in your posting. ***** Dear all, I have been asked to propose a protocol to fix Covid-19 infected cultured cells for fluorescence observations and for TEM embedding. I remembered to have seen Marks question (see below) and downloaded the PDF he is mentioning. Did anybody find and maybe tested in between additional protocols that could be recommended? Thanks a lot and best wishes, Christoph BIOIMAGING CENTER University of Geneva - Science II Room 245 30, Quai Ernest Ansermet CH - 1211 Genève 4 Dr. Christoph R. Bauer -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Mark Sanders Sent: 16 April 2020 19:03 To: [hidden email] Subject: Tissue fixations for safe and functional COVID-19 imaging ***** To join, leave or search the confocal microscopy listserv, go to: https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=02%7C01%7Checkman%40bgsu.edu%7C0c57ad04507c4a1c4ce308d7fbff3fda%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254946012403578&sdata=d3ksxZOdeifdtPiwsomRCCDA9xURlPOkNz27nz0m0dQ%3D&reserved=0 Post images on https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=02%7C01%7Checkman%40bgsu.edu%7C0c57ad04507c4a1c4ce308d7fbff3fda%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254946012403578&sdata=7An%2FhqOjsU24Ls6xotvDPeckQx4vey8VHbHZW%2FEvAUI%3D&reserved=0 and include the link in your posting. ***** Greetings Colleagues, I hope this email request finds you well. Many of our shared imaging resources may be requested to image COVID-19 infected tissues samples. Samples containing live virus are at least a BSL-3 situation so many labs must successfully fix the tissue to inactivate the virus to a manageable safety level while preserving the molecular integrity associated with the assay prior to the imaging steps. As is historically the case, there may not be a single fixation method for all assays, but they all must inactivate the virus for safe downstream steps. There are many options out there for viral inactivation methods for biological samples, including this one from WHO concerning the ebola virus ( https://nam02.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.paho.org%2Fhq%2Fdmdocuments%2F2014%2F2014-cha-procedures-inactivation-ebola.pdf&data=02%7C01%7Checkman%40bgsu.edu%7C0c57ad04507c4a1c4ce308d7fbff3fda%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254946012403578&sdata=WgRALBbP7OEaxDgyVF2E32ye21TawDXR%2F5ApKIrNMNo%3D&reserved=0 ) Has anyone come across preferred infected tissue fixation methods for COVID-19 samples for either FFPE or fixed cryosectioned tissues? References mandatory. Perhaps many of us will find the answer useful to either safely perform assays or to not do them at all based on our circumstances. Stay healthy, Mark *Working Remotely M-F as needed, (available via email)* -- Mark Sanders University of Minnesota Program Director Twin Cities Campus University Imaging Centers Nikon Center of Excellence <https://nam02.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.nikoninstruments.com%2FImaging-Centers%2FNikon-Centers-of-Excellence%2FNikon-CofE-Locations%2FAmericas%2FUniversity-of-Minnesota&data=02%7C01%7Checkman%40bgsu.edu%7C0c57ad04507c4a1c4ce308d7fbff3fda%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254946012403578&sdata=QfgR6kpT2tlxNf0Q3FadzcOvmN2VpK8FE9Zdocnxllw%3D&reserved=0> uic.umn.edu |
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Christoph Ruediger Bauer |
Re: Tissue fixations for safe and functional COVID-19 imaging
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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 Carol, Thanks for your feedback and to make it clear they can and want to split the samples for either immuno-fluorescence imaging or for TEM structure and treat them accordingly. So I need actually two protocols. Thanks, Christoph -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Carol Heckman Sent: 19 May 2020 16:59 To: [hidden email] Subject: Re: Tissue fixations for safe and functional COVID-19 imaging ***** 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. ***** That's a demanding task. To conserve antigenicity, you cannot fix with glutaraldehyde, at least not much glutaraldehyde, and to get good morphology you almost have to have some glutaraldehyde. Antigens vary in their degree of resistance to glutaraldehyde-induced inactivation. If you could split the sample in two, and process one half for immunofluorescence and the other for TEM, that would work best. Carol Heckman Bowling Green State University ________________________________ From: Confocal Microscopy List <[hidden email]> on behalf of Christoph Ruediger Bauer <[hidden email]> Sent: Tuesday, May 19, 2020 10:15 AM To: [hidden email] <[hidden email]> Subject: [EXTERNAL] Re: Tissue fixations for safe and functional COVID-19 imaging ***** To join, leave or search the confocal microscopy listserv, go to: https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=02%7C01%7Checkman%40bgsu.edu%7C0c57ad04507c4a1c4ce308d7fbff3fda%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254946012393579&sdata=AUOl7fqO26QSreknKotLJptmaEUrsk70wppcSNTsICs%3D&reserved=0 Post images on https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=02%7C01%7Checkman%40bgsu.edu%7C0c57ad04507c4a1c4ce308d7fbff3fda%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254946012393579&sdata=8S7c8RtZ8axAyZQvoCrVanS01Qk%2BmUQwxj8uaxwhsAk%3D&reserved=0 and include the link in your posting. ***** Dear all, I have been asked to propose a protocol to fix Covid-19 infected cultured cells for fluorescence observations and for TEM embedding. I remembered to have seen Marks question (see below) and downloaded the PDF he is mentioning. Did anybody find and maybe tested in between additional protocols that could be recommended? Thanks a lot and best wishes, Christoph BIOIMAGING CENTER University of Geneva - Science II Room 245 30, Quai Ernest Ansermet CH - 1211 Genève 4 Dr. Christoph R. Bauer -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Mark Sanders Sent: 16 April 2020 19:03 To: [hidden email] Subject: Tissue fixations for safe and functional COVID-19 imaging ***** To join, leave or search the confocal microscopy listserv, go to: https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=02%7C01%7Checkman%40bgsu.edu%7C0c57ad04507c4a1c4ce308d7fbff3fda%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254946012403578&sdata=d3ksxZOdeifdtPiwsomRCCDA9xURlPOkNz27nz0m0dQ%3D&reserved=0 Post images on https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=02%7C01%7Checkman%40bgsu.edu%7C0c57ad04507c4a1c4ce308d7fbff3fda%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254946012403578&sdata=7An%2FhqOjsU24Ls6xotvDPeckQx4vey8VHbHZW%2FEvAUI%3D&reserved=0 and include the link in your posting. ***** Greetings Colleagues, I hope this email request finds you well. Many of our shared imaging resources may be requested to image COVID-19 infected tissues samples. Samples containing live virus are at least a BSL-3 situation so many labs must successfully fix the tissue to inactivate the virus to a manageable safety level while preserving the molecular integrity associated with the assay prior to the imaging steps. As is historically the case, there may not be a single fixation method for all assays, but they all must inactivate the virus for safe downstream steps. There are many options out there for viral inactivation methods for biological samples, including this one from WHO concerning the ebola virus ( https://nam02.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.paho.org%2Fhq%2Fdmdocuments%2F2014%2F2014-cha-procedures-inactivation-ebola.pdf&data=02%7C01%7Checkman%40bgsu.edu%7C0c57ad04507c4a1c4ce308d7fbff3fda%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254946012403578&sdata=WgRALBbP7OEaxDgyVF2E32ye21TawDXR%2F5ApKIrNMNo%3D&reserved=0 ) Has anyone come across preferred infected tissue fixation methods for COVID-19 samples for either FFPE or fixed cryosectioned tissues? References mandatory. Perhaps many of us will find the answer useful to either safely perform assays or to not do them at all based on our circumstances. Stay healthy, Mark *Working Remotely M-F as needed, (available via email)* -- Mark Sanders University of Minnesota Program Director Twin Cities Campus University Imaging Centers Nikon Center of Excellence <https://nam02.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.nikoninstruments.com%2FImaging-Centers%2FNikon-Centers-of-Excellence%2FNikon-CofE-Locations%2FAmericas%2FUniversity-of-Minnesota&data=02%7C01%7Checkman%40bgsu.edu%7C0c57ad04507c4a1c4ce308d7fbff3fda%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254946012403578&sdata=QfgR6kpT2tlxNf0Q3FadzcOvmN2VpK8FE9Zdocnxllw%3D&reserved=0> uic.umn.edu |
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Carol Heckman |
Re: Tissue fixations for safe and functional COVID-19 imaging
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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, Christophe, We published a protocol for TEM on cultured cells in Nature Protocols. If you just search those terms with my name, Heckman, the Google should bring it up. For fluorescence, we fix briefly (10 min) with 3% formaldehyde made fresh from paraformaldehyde in cytoskeletal buffer. Rinse exhaustively before putting in the antibodies. Carol Heckman ________________________________ From: Confocal Microscopy List <[hidden email]> on behalf of Christoph Ruediger Bauer <[hidden email]> Sent: Tuesday, May 19, 2020 11:05 AM To: [hidden email] <[hidden email]> Subject: [EXTERNAL] Re: Tissue fixations for safe and functional COVID-19 imaging ***** To join, leave or search the confocal microscopy listserv, go to: https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=02%7C01%7Checkman%40bgsu.edu%7Ce4dc51cde21a48c0e91408d7fc062cb6%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254975758601842&sdata=Tr0JWpYvJdTP2nodKIWasAsjzw%2B2VjExUAyQSaH%2FtS0%3D&reserved=0 Post images on https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=02%7C01%7Checkman%40bgsu.edu%7Ce4dc51cde21a48c0e91408d7fc062cb6%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254975758601842&sdata=d7zCEA5I4PrFzZev3wjGSz9giCtaCjI7xh5BsP%2B0pA8%3D&reserved=0 and include the link in your posting. ***** Hi Carol, Thanks for your feedback and to make it clear they can and want to split the samples for either immuno-fluorescence imaging or for TEM structure and treat them accordingly. So I need actually two protocols. Thanks, Christoph -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Carol Heckman Sent: 19 May 2020 16:59 To: [hidden email] Subject: Re: Tissue fixations for safe and functional COVID-19 imaging ***** To join, leave or search the confocal microscopy listserv, go to: https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=02%7C01%7Checkman%40bgsu.edu%7Ce4dc51cde21a48c0e91408d7fc062cb6%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254975758601842&sdata=Tr0JWpYvJdTP2nodKIWasAsjzw%2B2VjExUAyQSaH%2FtS0%3D&reserved=0 Post images on https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=02%7C01%7Checkman%40bgsu.edu%7Ce4dc51cde21a48c0e91408d7fc062cb6%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254975758601842&sdata=d7zCEA5I4PrFzZev3wjGSz9giCtaCjI7xh5BsP%2B0pA8%3D&reserved=0 and include the link in your posting. ***** That's a demanding task. To conserve antigenicity, you cannot fix with glutaraldehyde, at least not much glutaraldehyde, and to get good morphology you almost have to have some glutaraldehyde. Antigens vary in their degree of resistance to glutaraldehyde-induced inactivation. If you could split the sample in two, and process one half for immunofluorescence and the other for TEM, that would work best. Carol Heckman Bowling Green State University ________________________________ From: Confocal Microscopy List <[hidden email]> on behalf of Christoph Ruediger Bauer <[hidden email]> Sent: Tuesday, May 19, 2020 10:15 AM To: [hidden email] <[hidden email]> Subject: [EXTERNAL] Re: Tissue fixations for safe and functional COVID-19 imaging ***** To join, leave or search the confocal microscopy listserv, go to: https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=02%7C01%7Checkman%40bgsu.edu%7Ce4dc51cde21a48c0e91408d7fc062cb6%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254975758601842&sdata=Tr0JWpYvJdTP2nodKIWasAsjzw%2B2VjExUAyQSaH%2FtS0%3D&reserved=0 Post images on https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=02%7C01%7Checkman%40bgsu.edu%7Ce4dc51cde21a48c0e91408d7fc062cb6%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254975758601842&sdata=d7zCEA5I4PrFzZev3wjGSz9giCtaCjI7xh5BsP%2B0pA8%3D&reserved=0 and include the link in your posting. ***** Dear all, I have been asked to propose a protocol to fix Covid-19 infected cultured cells for fluorescence observations and for TEM embedding. I remembered to have seen Marks question (see below) and downloaded the PDF he is mentioning. Did anybody find and maybe tested in between additional protocols that could be recommended? Thanks a lot and best wishes, Christoph BIOIMAGING CENTER University of Geneva - Science II Room 245 30, Quai Ernest Ansermet CH - 1211 Genève 4 Dr. Christoph R. Bauer -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Mark Sanders Sent: 16 April 2020 19:03 To: [hidden email] Subject: Tissue fixations for safe and functional COVID-19 imaging ***** To join, leave or search the confocal microscopy listserv, go to: https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=02%7C01%7Checkman%40bgsu.edu%7Ce4dc51cde21a48c0e91408d7fc062cb6%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254975758601842&sdata=Tr0JWpYvJdTP2nodKIWasAsjzw%2B2VjExUAyQSaH%2FtS0%3D&reserved=0 Post images on https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=02%7C01%7Checkman%40bgsu.edu%7Ce4dc51cde21a48c0e91408d7fc062cb6%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254975758601842&sdata=d7zCEA5I4PrFzZev3wjGSz9giCtaCjI7xh5BsP%2B0pA8%3D&reserved=0 and include the link in your posting. ***** Greetings Colleagues, I hope this email request finds you well. Many of our shared imaging resources may be requested to image COVID-19 infected tissues samples. Samples containing live virus are at least a BSL-3 situation so many labs must successfully fix the tissue to inactivate the virus to a manageable safety level while preserving the molecular integrity associated with the assay prior to the imaging steps. As is historically the case, there may not be a single fixation method for all assays, but they all must inactivate the virus for safe downstream steps. There are many options out there for viral inactivation methods for biological samples, including this one from WHO concerning the ebola virus ( https://nam02.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.paho.org%2Fhq%2Fdmdocuments%2F2014%2F2014-cha-procedures-inactivation-ebola.pdf&data=02%7C01%7Checkman%40bgsu.edu%7Ce4dc51cde21a48c0e91408d7fc062cb6%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254975758601842&sdata=KI%2FujVNTTGtmEKSlvkoS9Oe89MVeH%2B8A0fyWrmhcd1I%3D&reserved=0 ) Has anyone come across preferred infected tissue fixation methods for COVID-19 samples for either FFPE or fixed cryosectioned tissues? References mandatory. Perhaps many of us will find the answer useful to either safely perform assays or to not do them at all based on our circumstances. Stay healthy, Mark *Working Remotely M-F as needed, (available via email)* -- Mark Sanders University of Minnesota Program Director Twin Cities Campus University Imaging Centers Nikon Center of Excellence <https://nam02.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.nikoninstruments.com%2FImaging-Centers%2FNikon-Centers-of-Excellence%2FNikon-CofE-Locations%2FAmericas%2FUniversity-of-Minnesota&data=02%7C01%7Checkman%40bgsu.edu%7Ce4dc51cde21a48c0e91408d7fc062cb6%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C1%7C637254975758611799&sdata=scGBnWB4UsNdJJVXvsQ9YMSvoClw6xH0VCUsd5CRcMQ%3D&reserved=0> uic.umn.edu |
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