digesting thin sections

BIOHEAD GELLMORE at PEARL.TUFTS.EDU
Fri Jun 19 15:56:00 EST 1992


Sandra, are your thin sections in plastic or in paraffin? For our lab,
sections in Spurrs resin, and those in paraffin (7-9 um thick) have
stuck well to slides previously coated in alcoholic gelatin. We clean
slides in acidified 70% EtOH (3 drops 1N HCl/ 50 ml of 70% EtOH), by dipping
them in the stuff, and rubbing each slide off with lint-free cloth. Thebn
then, the pre-cleaned slides are immersed for 3-5 min (agitate) in
alcoholic gelatin, then dried in a 60 degree C oven for 2-12 hrs. Sections
stick real well to these slides which can be kept in their dry coated
condition indefinitely (stored in a slide box). We digest sections with
proteinases, as well as other enzymes, and we run fairly turbulent
immunohistochem on sections too. The subbing seems to hold.

So, what is alcoholic gelatin? : Heat 125 ml of Dist water to 50 degrees C
            -gradually add 1.5 gr of gelatin, and stir till dissolved (2-3 min)
            -remove from heat and add 100 ml Dist water
                                  and 285 ml 70% EtOH
            -mix, and store at room temp.

Hope this helps! Several labs have stopped by to use this from us.

Ref: Bloom, F.E., and ELF Battenberg. 1989. Imunocytochemistry of endorphins
and enkephalins. pp 679-696 IN Neuroendocrine peptide methodology. Academic
Press.



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