Re: [Histonet] querry

From:Gayle Callis

I only use chrome gelatin for decalcified bone sections IF the plus charge 
(Silane coated) slides DO NOT work.  You can make it up with different 
bloom gelatins (bloom indicating size of gelatin molecule 100 small, 275 
and 300 large).  The reason I do not care for gelatin, is background 
staining with hematoxylin, can be reduced by dipping presubbed slides in 
NBF 10 dips, rinsing well and storing in cool dry box.

DO NOT COAT SILANE  aka PLUS CHARGE SLIDES, it is an extra expense and the 
gelatin (protein) goes over the top of the Plus coating, rendering it 
useless.  This is clearly stated in package inserts with Plus Charge slides.

5gm gelatin in 1 liter distilled water, add 1 gm chromium potassium 
sulfate, dip clean microscope glass slides in this,
air dry and store in a cool dry place. You can these slides like regular 
slide to pick up section and do not add anything to waterbath, additional 
adhesives will cause more unsightly background.


Add 10 mls of this stock subbing solution to a 2 liter waterbath, like 
adding commerically available adhesives
to a waterbath, dry sections in normal way - We dry bone sections FLAT at 
37-40C for a couple of days, overnight is too minimal.  Soft tissues can be 
dried in an oven.

Chromium is considered very toxic, so use precautions when preparing the 
subbing solution.  We preferred the waterbath method when doing a large 
number of blocks.  Lessens preparation time to just have it in a waterbath 
already. We only use this subbing solution with soft tissue as a backup 
when all else fails to keep sections on a slide.  We never use it for 
immunstaining purposes.
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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