Re: [Histonet] Frozen sections and cold acetone
I have always "fixed" in RT acetone for 10 mins Have compared 0-60 mins/4C to RT acetone Lower temps just limit the rate of reaction, imho. I note "nuclear streaming" when I use acetone at any temp/time. Imho, acetone is not an effective fixativeit's a delipidiser. So, give it 10 mins at RT. If anyone thinks it's effective because it is a dehydrant….well, one re- hydrates the section to carry out IHC/ICC/IF. It works very well for only a few abs. When testing new abs out on FS/cell monolayers ( unfixed) I always compare Formalin, Acetone, Methanol and methanol/acetone 1:1 Imho Best wishes to a great site/membership. Carl ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Frozen sections and cold acetone...
Cold acetone, and cold ethanol, were used to fix tissues because they left enzymes unaffected and still demonstrable. This was in the early days of enzyme histochemistry. Pearse' Histochemistry: Theoretical and applied,3rd edition, volume 1, page 85 discusses it. I could send a scan if you wanted. Bryan Llewellyn Morken, Timothy via Histonet wrote: Can anyone give me a rational for using cold (refrig or freezer-temp) acetone to fix frozen sections? Or a rational for using RT acetone. This is for kidney or muscle bx frozens for immmunofluroescence or immunoperoxidase staining. Normally they air dry for at least 15 minutes (just waiting for frozen sectioning to be completed) before going into acetone. Just wondering if we can reduce complexity... I haven't seen anything saying why cold acetone is used, just instructions to do so. I always wonder about such things... Tim Morken Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Center ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Frozen sections and cold acetone...
Can anyone give me a rational for using cold (refrig or freezer-temp) acetone to fix frozen sections? Or a rational for using RT acetone. This is for kidney or muscle bx frozens for immmunofluroescence or immunoperoxidase staining. Normally they air dry for at least 15 minutes (just waiting for frozen sectioning to be completed) before going into acetone. Just wondering if we can reduce complexity... I haven't seen anything saying why cold acetone is used, just instructions to do so. I always wonder about such things... Tim Morken Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Center ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
