Re: Nuclear Staining

<< Previous Message | Next Message >>
From:amos brooks <>
Content-Type:text/plain; charset=us-ascii

    A while back, we had a problem like this. The problem is usually a ghost
problem, it is bad one day and just when you think you got rid of it it comes
back. We think it is a dehydration artefact. It is often worsened by soaking
blocks while cutting. Make sure the slides are adequately heated to remove all
water before staining.
    The main culprit probably goes as far back as processing. Make sure the
tissue is completely infiltrated. Good clean paraffin will remove the xylene.
Residual xylene will be present in the sections after cutting the blocks. When
the tissue is rehydrated during staining, the xylene remaining in the tissue
will obscure the nuclear staining and produce a hazy ring around the nuclei,
xylene + alcohol + water = precipitate. (...QED)
Amos Brooks

"SALA,ROBERT" wrote:

> Please forgive me if I am going over old discussions.
> We are having a problem with our nuclear staining.  It especially occurs on
> tiny biopsies and on lymphoid tissue.  It is that hazy washed out nuclear
> pattern.  We have tried about everything to correct the problem.  One our
> paths is adamant that it is a heat artifact, I am leaning to incomplete
> dehydration.  We have no heat above 63 C. in any of our procedure, we
> dehydrate in ethanol and clear in a xylene substitute called Clear-Rite III.
> This artifact in intermittent and does not affect all tissues on the run.
> It appears to get worse towards the bottom of the slides.  VERY FRUSTRATING!
> If anyone has any suggestions on how to correct this please let me know.
> Thanks everyone.
> Robert M. Sala M.T.(ASCP)
> Histology Supervisor
> Sioux Valley Hospital
> 1100 S. Euclid Ave.
> Sioux Falls, SD  57117-5039

<< Previous Message | Next Message >>