[Histonet] PFA Fixed Tissue not sticking to slides?

[jenny sandy]

Thank you Charles Scouton for responding.  The slides are positive charged slides from IMEB.

Thank you Elizabeth Chilpala for responding.  The treatment after sectioning is one I’ve forwarded to the service that is processing for us.  I only do the PFA/sucrose/embedding part.  My understanding is it is pretty standard (?) for cryo sections for H&E and IHC, but I am checking that out.

Jennifer Eveleth

Original message below:

Dear HistoFolks,

Has anyone had experience with cryo-sectioned tissue that is PFA fixed not adhering to slides?

The tissue is rabbit cornea culture, 4% PFA fixed O/N, then run through 20% sucrose gradient, then frozen in OCT.

We think that the PFA fix is much longer than needed for a tissue this thin, and were going to try 2 hr fix. Would over-fixation cause the section to not adhere?  

The PFA/Sucrose fixation really improves the morphology and the IHC stains that we have run so far.

Thank you in advance for your help.

Jennifer Eveleth