Methods and Techniques Discussion => Electron Microscopy => Topic started by: ssinha on August 10, 2004, 02:56:04 PM

Title: Tissue specific fixatives
Post by: ssinha on August 10, 2004, 02:56:04 PM
I am trying to characterize a zebrafish mutant and am having trouble with preservation of various tissues. I am currently using a cacodylate buffer for fixation and I was wondering if that fixative is good enough for all tissues. Would anyone be able to tell me if there are tissue specific fixatives that I may need to use?
Thank you   :D
Title: Tissue specific fixatives
Post by: immunoem on August 11, 2004, 03:13:40 AM
which fixative regime are you using and which tissues are you having problems with? Some fixatives are better with certain tissues. For instance, picric acid-based fixatives (such as Bouin's) work fine with most tissues but are not recommended for kidney. I would have thought that the standard PFA+glutaraldehyde fix would be a good general fixative. Are you sure that your problem is not due to slow tissue penetration by the fixative? I assume that you are immersion fixing, in which case some tissues may be fixed later than others. Also glutaraldehyde doesn't penetrate tissue very fast which also may affect the preservation of morphology.
I don't work with zebrafish, but is it possible to dissect out the tissues of interest and drop them into fixative (perhaps you already do this)? If you have access to a suitable microwave, you could try microwave-assisted fixation which should speed up the process. Finally, what do other people use for zebrafish?
Not sure if the above helps much, but that's my 2cts worth.
Good luck.