H&E problems

From:Andrea Voogt <ndrvgt@dordt.edu>

Hi all,
I've been staining plastic-embedded sections with H&E and running into a few problems.  First of all, on most of the slides the sections (which weren't wrinkled when I began) come out with lots of small wrinkles throughout the plastic - including the tissue.  I think this might have something to do with either the length of staining in eosin (I've been using 8 minutes) or the 1-second rinse in 95% EtOH, because I added 95% EtOH to my finished (but no good) slide and the wrinkling got worse.  However, I don't know how to correct the problem.
The other problem I've been having is that the eosin looks cracked on the tissue - sort of like dried earth.  This creates a problem in that the "cracks" are easily mistaken for cell membranes, creating artificial boundaries between the cells.  But I've noticed that the cracks often go right through nuclei, so they can't be the actual cell membranes.
I don't know if this makes a difference or not, but I've also noticed that my staining is turning out a lot darker than the last time I did it (a year ago).
Thanks in advance for all your help!!
Andrea Voogt
Dordt College   

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