RE: [Histonet] Glycerin-Gelatine bubble-phenomen

From:"Barry R Rittman"

I believe that part of the  problem is that you are drying the slides before adding the glycerin gelatin. 
I would recommend shaking excess water from the slide, placing the slide on a warming plare at 60 degrees C. Add a few drops of glycerin gelatin and leave for a few minutes to allow any bubbles to rise to the surface.
Then add coverslip.
The bubbles are probably coming from the glycerin as sections mounted just in glycerin always develop bubbles after some days.
Suggestion is to place the glycerin in a tube in an ultrasonic bath to get bubbles out before making the glycerin gelatin or simply doing the same to the molten glycerin gelatin.

	-----Original Message----- 
	From: on behalf of Barbara Bublava 
	Sent: Mon 9/20/2004 3:48 AM 
	Subject: [Histonet] Glycerin-Gelatine bubble-phenomen

	Hi Everybody!
	I have an interesting phenomen mounting slides with Glyceringelatine
	When I mount my slides they do not have bubbles. Oil Red O stains do not develop bubbles even after some time but when I mount CAE - Chloracetate Esterase (I mount in the same way I do the ORO) the slides develop bubbles after some hours and after some days the whole section is covered with bubbles, but there are no bubbles around the section!
	Why does this happen and how can I prevent this???
	I use Glyceringelatine at 60°C, slides out of dest. water, excess water shaked of. I did also try to dry the slides bevore mounting but there happend to be more bubbles from the beginning and it did not change this strange effect.
	Sometimes, when I am in a hurry I further hardening of the Glyceringelatine at fridge (4°C) but I can not see any positive or negative effect on bubbles doing that.
	Thanks to everyone
	Barbara, Vienna
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