Re: [Histonet] neutrophils

From:Barbara Bublava

Hi Dorothy

I did have a similar problem with some specimens lately.

So I did a little test with some human spleen:
one piece was properly fixed with formaldehyde, one piece of the same
spleen wasnīt fixed in formaldehyde but transferred directly to 70%
Ethanole, and one piece was improperly fixed in formaldehyde and then
transferred to 70% Ethanole.
All pieces were processed together after 24 hours.

The piece transferred directly to 70% Ethanol didnīt work at all, the
improperly fixed speciemen did stain irregularly - i could clearly see
which parts were fixed and which not. The correct fixated piece did work

I did not have time to make further tests to see if autolysis is the
problem or if it is the alcohol - or maybe the water in the alcohol?

best regards

Barbara Bublava
Medical University of Vienna

Webb, Dorothy L wrote:
> We are staining some research tissue (rabbitt?) for neutrophils using
> the chloroacetate esterase kit from Sigma with an inflamed appendix for
> a control.  The control turned out beautiful, but, nothing is showing up
> in the research tissue.  We did not  gross in the research tissue, only
> processed it according to our routine processing schedule.  According to
> the "fellow" who is doing this project, the formalin was expired on the
> research tissue.  Does anyone have any ideas on this case??  I am at a
> loss and the pathologist helping the "fellow" to read these tissues out
> is wanting me to come up with a solution, of course!!  So, HELP and
> thanks, as always!!
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