[Histonet] fixation for frozen tissue

[Rita Angel]

Hi all,

I have several questions about fixing frozen tissue. In the past, we've 
always frozen tissue fresh from the animal into frozen embedding media and 
then immersing into liquid nitrogen.

I have an investigator that brought frozen tissue that he first fixed in 
formalin, then froze. I'm unable to get sections because the tissue is 
soft. It seems there is still moisture in the tissue, so I melted the block 
& blotted off the tissue, then re-froze. I'm still not able to get sections 
although it doesn't seem as mushy now. It still seems like the tissue is 
wet.Does anyone have any suggestions?

He was also asking about a protocol for sucrose, and I was wondering if 
anyone could get one to me? Also why do people use this procedure, and when 
do you need to use it?

Thanks for all your help,

Rita Angel, HT