RE: [Histonet] neutrophils
We routinely use this kit for our Esterases, and it works well for us. The one thing you should tell your researcher, is that this kit is not specific to just neutrophils. It is specific to any cell in the granulocytic lineage, so your basophils, and eosinphils will also stain.
We have had some research material stain negatively (this was mouse in our case) so I wonder if this kit is geared to human enzymatic activity specifically. I would try calling Sigma and see it this is the case.
Otherwise, the insert states that there are a few things that can cause a false negative, although if your control works, I would say that it is something to do with the tissue itself (or the fixation). I don't think that expired formalin would do it as we get samples from Cameroon on a regular basis, and the formalin in them is definitely not great, and they work fine.
This may not help much, but I would try Sigma first and see if the kit has been designed for non-human tissue!
Best of Luck!
Tech II Special Stains
Calgary Laboratory Services
#9 - 3535 Research Road NW
[mailto:email@example.com]On Behalf Of Webb,
Sent: June 9, 2006 6:58 AM
Subject: [Histonet] neutrophils
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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