Help for immuno-histology needed

Xianze Chen xianze at McRCIM.McGill.EDU
Fri Jun 25 05:50:18 EST 1993


Hello all:

	I am doing some immunohistochemical studies of rat eyes for my Ph.D
project. I am using APAAP staining method.

	I need some help for preparing eye grozen sections of good qaulity. I am experiencing some problems of keeping getting collapsed eyes when I cut them.
I guess it is because frozen vitreous within the eye ball is not as strong as 
the frozen OCT (frozen medium) out of eye ball. Has any one got any experience 
in solving this problem?

	I have tried a method. I removed the cornea and soaked the eye
 in OCT/PBS, then OCT for one hour to let OCT penetrate into the eye. But in 
order to keep tissue fresh, I had to use 100% Ethanol perfussion fixation
before I remove the eye. Does this have any side effect at all?

 	Any comments will be highly appreciated.

Many thanks in advance.


Daniel Y-F Cheng, MB ChB	opt018 at aberdeen.ac.uk
Dept. of Ophthalmology		Tel: (0224) 681818-53782
Medical School
University of Aberdeen
Aberdeen AB9 2ZD
Scotland
U.K.



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