Sample Prep: Frozen sections not 'sticking' to slides
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Ellen T. Arena |
Sample Prep: Frozen sections not 'sticking' to slides
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hello all - I am actually having a sample-prep issue - I am hoping someone can help... I am wanting to mount 'thick' frozen sections (~30um) of tissue on SuperFrost slides for staining/imaging. I am finding the tissues come 'off' quite easily during the staining/washing procedure. I have used the typical methods with OCT-removal (with PBS), fixation with PFA, etc. I am not quite sure if any 'trick' exists for these thicker sections that I might be missing... ??? Please let me know if you have any advice. Many thanks. Ellen ----------------------------------------------------------------------------- Ellen T. Arena, PhD Pasteur Foundation Postdoctoral Fellow Unité de Pathogénie Microbienne Moléculaire INSERM U786 Institut Pasteur 28 rue du Dr Roux F - 75724 PARIS Cédex 15 France Tel: (33-0) 1 40 61 37 71 Fax: (33-0) 1 45 68 89 53 [hidden email] ----------------------------------------------------------------------------- |
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Dr. Judith Drazba |
Re: Sample Prep: Frozen sections not 'sticking' to slides
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi Ellen, We do this all the time but find that frozen sections, especially thick ones, only stick to Superfrost Plus slides. Regular Superfrost slides don't work. Whatever they do to make them + (plus) - makes all the difference. You can always tell which slides your using because the Plus slides have little + on the corners. Hope this helps. Judy -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Ellen T. Arena Sent: Tuesday, June 25, 2013 4:48 AM To: [hidden email] Subject: Sample Prep: Frozen sections not 'sticking' to slides ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hello all - I am actually having a sample-prep issue - I am hoping someone can help... I am wanting to mount 'thick' frozen sections (~30um) of tissue on SuperFrost slides for staining/imaging. I am finding the tissues come 'off' quite easily during the staining/washing procedure. I have used the typical methods with OCT-removal (with PBS), fixation with PFA, etc. I am not quite sure if any 'trick' exists for these thicker sections that I might be missing... ??? Please let me know if you have any advice. Many thanks. Ellen ----------------------------------------------------------------------------- Ellen T. Arena, PhD Pasteur Foundation Postdoctoral Fellow Unité de Pathogénie Microbienne Moléculaire INSERM U786 Institut Pasteur 28 rue du Dr Roux F - 75724 PARIS Cédex 15 France Tel: (33-0) 1 40 61 37 71 Fax: (33-0) 1 45 68 89 53 [hidden email] ----------------------------------------------------------------------------- =================================== Please consider the environment before printing this e-mail Cleveland Clinic is ranked one of the top hospitals in America by U.S.News & World Report (2012). Visit us online at http://www.clevelandclinic.org for a complete listing of our services, staff and locations. Confidentiality Note: This message is intended for use only by the individual or entity to which it is addressed and may contain information that is privileged, confidential, and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient or the employee or agent responsible for delivering the message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please contact the sender immediately and destroy the material in its entirety, whether electronic or hard copy. Thank you. |
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In reply to this post by Ellen T. Arena
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi Ellen, A few suggestions: -Use Super Frost Plus slides which has a charge to it and it will prevent the sections from falling off during IHC processing. -Poly-lysine coating or chrome alum-gelatin coating might do the trick -use silanized slides For us, the super frost plus slides worked well. I found these tricks on the IHC World website (http://www.ihcworld.com/_faq/histology-faq/section/s1.htm). Good luck. Sathya Srinivasan Manager RUN Microscopy Core Facility (www.ucalgary.ca/runcore) Room # 126 HMRB 3330 Hospital Drive, NW Calgary, AB T2N 4N1 Canada > Date: Tue, 25 Jun 2013 03:47:40 -0500 > From: [hidden email] > Subject: Sample Prep: Frozen sections not 'sticking' to slides > To: [hidden email] > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hello all - > > I am actually having a sample-prep issue - I am hoping someone can help... > > I am wanting to mount 'thick' frozen sections (~30um) of tissue on > SuperFrost slides for staining/imaging. I am finding the tissues come 'off' > quite easily during the staining/washing procedure. I have used the typical > methods with OCT-removal (with PBS), fixation with PFA, etc. I am not quite > sure if any 'trick' exists for these thicker sections that I might be > missing... ??? > > Please let me know if you have any advice. > > Many thanks. > > Ellen > > > ----------------------------------------------------------------------------- > Ellen T. Arena, PhD > Pasteur Foundation Postdoctoral Fellow > Unité de Pathogénie Microbienne Moléculaire > INSERM U786 > Institut Pasteur > 28 rue du Dr Roux > F - 75724 PARIS Cédex 15 > France > Tel: (33-0) 1 40 61 37 71 > Fax: (33-0) 1 45 68 89 53 > [hidden email] > ----------------------------------------------------------------------------- |
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Kilgore, Jason-2 |
Re: Sample Prep: Frozen sections not 'sticking' to slides
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In reply to this post by Dr. Judith Drazba
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** I agree with Judy. Additionally, if you use gelatin or poly-L-lysine slides, you can do an aldehyde fixative after mounting the section, which will crosslink in place the tissue with they lysines. Jason -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Drazba, Ph.D., Judy Sent: Tuesday, June 25, 2013 7:40 AM To: [hidden email] Subject: Re: Sample Prep: Frozen sections not 'sticking' to slides ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi Ellen, We do this all the time but find that frozen sections, especially thick ones, only stick to Superfrost Plus slides. Regular Superfrost slides don't work. Whatever they do to make them + (plus) - makes all the difference. You can always tell which slides your using because the Plus slides have little + on the corners. Hope this helps. Judy -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Ellen T. Arena Sent: Tuesday, June 25, 2013 4:48 AM To: [hidden email] Subject: Sample Prep: Frozen sections not 'sticking' to slides ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hello all - I am actually having a sample-prep issue - I am hoping someone can help... I am wanting to mount 'thick' frozen sections (~30um) of tissue on SuperFrost slides for staining/imaging. I am finding the tissues come 'off' quite easily during the staining/washing procedure. I have used the typical methods with OCT-removal (with PBS), fixation with PFA, etc. I am not quite sure if any 'trick' exists for these thicker sections that I might be missing... ??? Please let me know if you have any advice. Many thanks. Ellen ----------------------------------------------------------------------------- Ellen T. Arena, PhD Pasteur Foundation Postdoctoral Fellow Unité de Pathogénie Microbienne Moléculaire INSERM U786 Institut Pasteur 28 rue du Dr Roux F - 75724 PARIS Cédex 15 France Tel: (33-0) 1 40 61 37 71 Fax: (33-0) 1 45 68 89 53 [hidden email] ----------------------------------------------------------------------------- =================================== Please consider the environment before printing this e-mail Cleveland Clinic is ranked one of the top hospitals in America by U.S.News & World Report (2012). Visit us online at http://www.clevelandclinic.org for a complete listing of our services, staff and locations. Confidentiality Note: This message is intended for use only by the individual or entity to which it is addressed and may contain information that is privileged, confidential, and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient or the employee or agent responsible for delivering the message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please contact the sender immediately and destroy the material in its entirety, whether electronic or hard copy. Thank you. |
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SUBSCRIBE CONFOCALMICROSCOPY Sara Smith |
Re: Sample Prep: Frozen sections not 'sticking' to slides
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Our histology core also find that Super Frost Plus slide works best with thick sections! Good luck! Yi On Tue, Jun 25, 2013 at 4:29 PM, Kilgore, Jason <[hidden email]>wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > I agree with Judy. Additionally, if you use gelatin or poly-L-lysine > slides, you can do an aldehyde fixative after mounting the section, which > will crosslink in place the tissue with they lysines. > > Jason > > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] > On Behalf Of Drazba, Ph.D., Judy > Sent: Tuesday, June 25, 2013 7:40 AM > To: [hidden email] > Subject: Re: Sample Prep: Frozen sections not 'sticking' to slides > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hi Ellen, > > We do this all the time but find that frozen sections, especially thick > ones, only stick to Superfrost Plus slides. Regular Superfrost slides don't > work. Whatever they do to make them + (plus) - makes all the difference. > You can always tell which slides your using because the Plus slides have > little + on the corners. > > Hope this helps. > Judy > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] > On Behalf Of Ellen T. Arena > Sent: Tuesday, June 25, 2013 4:48 AM > To: [hidden email] > Subject: Sample Prep: Frozen sections not 'sticking' to slides > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hello all - > > I am actually having a sample-prep issue - I am hoping someone can help... > > I am wanting to mount 'thick' frozen sections (~30um) of tissue on > SuperFrost slides for staining/imaging. I am finding the tissues come 'off' > quite easily during the staining/washing procedure. I have used the > typical methods with OCT-removal (with PBS), fixation with PFA, etc. I am > not quite sure if any 'trick' exists for these thicker sections that I > might be missing... ??? > > Please let me know if you have any advice. > > Many thanks. > > Ellen > > > > ----------------------------------------------------------------------------- > Ellen T. Arena, PhD > Pasteur Foundation Postdoctoral Fellow > Unité de Pathogénie Microbienne Moléculaire INSERM U786 Institut Pasteur > 28 rue du Dr Roux > F - 75724 PARIS Cédex 15 > France > Tel: (33-0) 1 40 61 37 71 > Fax: (33-0) 1 45 68 89 53 > [hidden email] > > ----------------------------------------------------------------------------- > > =================================== > > > Please consider the environment before printing this e-mail > > Cleveland Clinic is ranked one of the top hospitals in America by U.S.News > & World Report (2012). > Visit us online at http://www.clevelandclinic.org for a complete listing > of our services, staff and locations. > > > Confidentiality Note: This message is intended for use only by the > individual or entity to which it is addressed and may contain information > that is privileged, confidential, and exempt from disclosure under > applicable law. If the reader of this message is not the intended > recipient or the employee or agent responsible for delivering the message > to the intended recipient, you are hereby notified that any dissemination, > distribution or copying of this communication is strictly prohibited. If > you have received this communication in error, please contact the sender > immediately and destroy the material in its entirety, whether electronic or > hard copy. > > Thank you. > |
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