Re: [Histonet] 1% gelatin/40% etOH mounting solution

From:Gayle Callis


I have never heard of using alcohol with gelatin to make up a section 
adhesive.   If you see ppt or gelatin coming out of solution, I would 
strongly suspect the gelatin is being "fixed" by the alcohol since gelatin 
is a protein and alcohol fixes proteins, what you see is probably 
precipitated (denatured) gelatin, not voodoo, just chemistry.  Just an 
educated guess here.

Chrome gelatin subbing solution:

Dissolve 10 g gelatin in 1 liter WARM distilled water, add 1 gm chromium
potassium sulfate, stire until dissolved, add a thymol crystal for 
preservation.   Final
concentration of gelatin is 1%.  You can also make this a 0.5% solution if
you wish.

Use 100 bloom  gelatin for most tissures.  For bone, use 275 bloom gelatin 
made from pig collagen.

Add 10 mls of this solution to 2 liter water bath or sub clean, uncoated 
microscope slides.

Reduce background staining by dipping dry subbed slides in NBF 10 times, 
rinse well, air dry, store.

Do NOT coat Plus Charge slides as these slides are already coated with 
silane to attract negatively charged tissue protein components.

At 11:22 AM 9/27/2005, you wrote:
>I am trying to get fixed tissue sections to stick onto slides better. I have
>tried subbed slides, plus slides (subbed and un-subbed) with only mediocre
>results. I was recently given a recipe for a 1% gelatin in 40% etOH mounting
>solution. This seems to work wonderfully on un-subbed plus slides, but I am
>having much difficulty in keeping the gelatin in solution long enough for it
>to be useful for mounting my sections. The recipe consists of heating
>distilled water (not over 60C) dissolving gelatin (enough for a 1% final
>concentration) and adding 80% etOH in two steps to the cooling gelatin
>solution, half soon after gelatin goes into solution and the other half
>sometime into the cooling of the solution. The recipe is vague since there
>is probably some voodoo that goes along with it. I have tried about 10
>different variations on this recipe, none work consistently. Does anyone
>have some tips, pointers or advice to help me out, or even another recipe
>that may work better. Thanks in advance!
>Brandon Munk
>Dept. Zoology and Physiology
>University of Wyoming
>Histonet mailing list

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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