[Histonet] help: holes in the spleen...


Hello, I am trying to get confocal images of spleens of Listeria infected mice.
Sadly, I have ran into a problem. I get some large holes in the tissue right
after cutting the block. These holes appear in areas where the spleen appears
almost black on the face of the block.The protocol I am following is the next
one:Isolate the spleen, incubate 4 to 6 hrs in Paraformaldehyde 1%, and leave
overnight in sucrose 30%. Spleens are included in OCT and frozen on dry ice.
All suggestions are welcomed.

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