[Histonet] massons trichrome

[Vinnie Della Speranza]

Rita,
jt is difficult to give you anything better than a guess without seeing what the stain looks like. However, it is entirely possible that what may appear to be aniline blue in the muscle cells is actually weigert's hematoxylin. which can remain if the water washes following weigerts hematoxylin staining are insufficient to remove excess stain from non-nuclear tissues. 

I like to  rinse weigert's stained sections in 50- 70% ethanol (8-10 dips in a coplin jar) following the wiegert's stain which is then followed by the water rinse in order to achieve a more thorough removal of the excess stain. You will note that the working weigerts solution  contains approx 50% alcohol and will more readily come out in a dilute alcohol than water alone.

Vinnie


Vinnie Della Speranza
Manager for Anatomic Pathology Services
Medical University of South Carolina
165 Ashley Avenue  Suite 309
Charleston, SC 29425
Ph: 843-792-6353
fax: 843-792-8974

>>> Rita Angel <RITA.ANGEL <@t> UC.EDU> 02/12/04 03:19PM >>>
hello all,

I was hoping someone could help us with a trichrome problem. We are 
suddenly having problems with the muscle in heart tissue staining purple 
instead of red. Our skin control is staining the same way. There are 2 
techs in the lab, and we're both having problems. The problems are with 
complete batches, not slides within the batch. We each stained 2 batches in 
a row, one batch stained with the purple muscle, and the 2nd batch with red 
muscle. We're leaving the slides in the solutions the same amount of time 
and rinsing the same amount of times, etc. We're using the same solutions 
from the same bottles, etc. (we make all of our own solutions). We are 
unable to think of what could be causing the problem. Also, our slides are 
still really black when they are finished rinsing after the wiegert's step. 
Could this be causing it? I appreciate any advice.

Thanks,

Rita Angel, HT
University of Cincinnati


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