To anyone who performs a NF-LFB (+PAS) combo stain using a Ventana
Benchmark:
I have been trying to stain a nerve (cross and longitudinal section) and
it keeps falling off of the slide after the neurofilament stain. It's
been cut onto a charged slide and has been baked 20 minutes prior to IHC
staining. After IHC staining, we have rinsed in Dawn-water per
protocol; by then the longitudinal section is peeling off the slide. I
tried skipping the Dawn rinse and have gone to straight 95% alcohol and
1 of 4 sections survived on the slide (others fell off).
Tissue problem - the nerve is well myelinated and fatty. If I drop the
water bath temp too low, the nerve doesn't flatten well; if the temp is
too high, it's shredding and spreading.
Has anyone out there had this problem and resolved it? Or do you have a
protocol that might have avoided it completely?
Any suggestions would be appreciated.
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