[Histonet] PFA Fixed tissue not sticking to slides?

[Due, Brice]

Hello Jennifer, yes over-fixation will cause adhesion problems. This applies to formalin fixed frozens as well as PFA fixed frozens.

I have had good luck using chromated gelatin to coat slides. I have successfully cut and stained lung specimens left in NBF for several months. Commercially, chromated gelatin is sold as STA-ON. I believe Leica now owns it, but you can make your own quite easily.

I tend to use STA-ON full strength, or minimally diluted. Spread some on a slide with a transfer pipette, taking care not to allow it to run onto the back of the slide. Shake off the excess and set vertical to dry. The coating MUST be dry before you use the slide to pick up sections. Freshly made slides work best.

After pick up, it is important to air-dry the section completely before immersing it in any solution. The moisture content of the frozen section must evaporate or the chromated gelatin will not hold the tissue. Don't worry, fixed sections do not suffer from artifacts when air-dried because the morphology is already locked by the fixation.

Best luck!
-brice






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