[Histonet] Re: MMA sections falling off

Gayle Callis gcallis <@t> montana.edu
Wed Dec 21 09:14:11 CST 2005

To increase the holding power, the gelatin can be 275 bloom, a larger 
gelatin molecule made from pig collagen.  This subbing solution suggested 
by Nancy is tried and true, also works for the nasty decalcified bone 
sections, generally larger ones. However, if the subbing solution causes 
too much blue background staining from hematoxylin, one can dip the subbed 
slides in NBF a couple of times, rinse well, dry and store in cool, dry 
place.  The NBF crosslinks the gelatin a bit and reduces blue background.

At 05:20 AM 12/21/2005, you wrote:
>We use Chrome-alum gel coated slides and prepare them as 
>follows:   dissolve 9 g gelatin(Fisher #G8-500) per 1000 ml distilled 
>water (solution A) while gently heating solution.  Dissolve 4 g Chromium 
>Potassium Sulfate (Fisher #C337-500) in distilled water (solution 
>B).  Prepare gelatin solution for slide dip:  Mix 500 ml Solution a with 
>19.25 ml solution B.  Heat to 70-75 Deg. C, dip slides 2 -4 minutes in 
>this solution.  Allow to dry vertically, then put in hot oven overnight 
>(60 deg. C).  To adhere sections to slides we place the 5 micron MMA 
>sections on slide, spread with 70% ethanol, cover with a piece of plastic 
>and stack slides then clamp with an office clamp.  Put slides in 37deg C 
>oven for two nights or for one night in 60 deg C oven (don't do the hot 
>oven if you are subsequently staining for enzymes).  For immuno we mount 
>our 5 micron plastic sections on hybridization slides from Scientific 
>Device Laboratory, Cat. #063 and place in hot oven 60 deg C 
>overnite.  Hope this helps!
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Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)

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