[Histonet] Re: H&E Stain

Johnson, Teri TJJ <@t> stowers.org
Thu Jan 20 10:00:27 CST 2011

Allison Scott wrote:

Hello to all in histoland and Happy New Year.  We are having issues with
our H&E stain.  The nuclei are staining very blue to purple and the
mucin is staining blue to purple-blue.  It is difficult to see the
nuclear detail.  The mucin is obscuring things.  We have not changed our
process for staining or processing.  The funny thing is that it is only
in the Biopsy cases, and it is every few slides.  The surgical  cases
are all right.  We checked the alcohol and xylene for water, and there
is not any.  My tech changed out the stain and we are staining a new
batch of slides.  If anyone has any idea what is wrong, any help would
be greatly appreciated.  I have gone over our processes and nothing has
changed.  The reagents are the same, the staining times are the same,
and the processing times are the same.  We are using the Shandon Gemini
stainer and VIP processor.

Allison Scott HT(ASCP)
Histology Supervisor
LBJ Hospital
Houston, Texas

When you say it is every few slides, is there variation within multiple slides of one case? If all the slides (levels) of a case look the same, I would suspect a problem with that particular case, not the processing and not the staining.
Are these all GI biopsies? Or do you see this in other needle biopsies, cervical biopsies and the like? GI biopsies tend to show hazy nuclei in epithelial cells commonly. There are several explanations and mostly are attributed to inadequate fixation.

It could be you a client who has changed how they collect and fix the specimens. Are they letting them dry on a gauze before putting them in fixative? It could be you have something out of place on the tissue processor. It could be there is water under the tissue on the slide and it is subject to high heat (microwave or oven) and is cooking the cells prior to staining. As pointed out previously, it could be a problem with your hematoxylin pH.

Good luck, and please let us know what the fix for this is when you get it figured out!

Best wishes,
Teri Johnson, HT(ASCP)QIHC
Head, Histology and Electron Microscopy
Stowers Institute for Medical Research
Kansas City, MO

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