[Histonet] Fixation and staining problems

From:Christine Tambasco

I have been having problems with tissue fixation and/or staining. I know one can really affect the other. I have been running the same program for 20 years. I had a Pm done on my tissue processor and completely changed all reagents. The stain is not crisp and some tissue appears raw during embedding. I process all types of tissues in the same program as I had for years. We are a small 120 bed hospital and only process one run a night up to 14o blocks. We are presently using Richard Allen 10% neutral buffered formalin v/v, grades of absolute ethyl alcohol we dilute to 70%, 80% and 95% from Pharmco, Formula 83 and paraffin type 9 from Richard Allen. We also use Richard Allen Hematoxylin 7211, and Richard Allen Eosin-y Alcoholic. Everything has been the same for at least a couple years. Am I missing something? The nuclei are pale with no chromatin.(The pathologist calls it "muddy"). And why are we getting unprocessed tissue? Especially the big (prostate,uterus) and fatty type tissue. Any ideas would be greatly appreciated. I am checking with all manufacturers on the lot numbers. So far nothing....
Thanks again.
Christine Tambasco, HT (ASCP)
St. Mary's Hospital 
Amsterdam, New York
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