[Histonet] querry

[Monfils, Paul]

Making up your own chrome gelatin is a bit tedious, though I did so for
years before commercial products became available.  Surgipath sells a chrome
gelatin product called STA-ON, which I have used for several years now.  A
small amount of the solution added to the water bath works very well for
many kinds of hard tissue, as well as necrotic tissue, blood clots, and
other materials which tend to detach from the slide.  It also works well on
sections of polymers and other foreign substances, which the average
clinical lab doesn't deal with very much, but which we have to section
frequently since we are a core research facility.  "Plus slides" don't work
on such materials, since these materials usually do not possess the net
negative charge that most tissues have.

> ----------
> From: 	histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Gayle
> Callis
> Sent: 	Thursday, September 29, 2005 8:10 AM
> To: 	Arvind Pundir; Histonet <@t> lists.utsouthwestern.edu
> Subject: 	Re: [Histonet] querry
> 
> 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.
> 
> OR
> 
> 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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