[Histonet] inconsistent H&E staining

[Geoff]

Failure to completely remove paraffin is often a cause of uneven 
staining. Change all of the xylenes and add another xylene.

Geoff

On 4/1/2015 12:06 PM, Julie Cohen wrote:
> Hi,
>
> I made slides of paraffin-embedded mouse small intestine (Swiss rolls), and stained them with Hematoxylin and Eosin.  Parts of the tissue on the same slide are stained dark with good structure.  Other areas look washed out with poor structure.  We realize that some of this could be caused by the orientation/structures captured, but similar tissue type looks paler as well.
>
> Has anyone had a similar experience, and could suggest an explanation for me to give to our client?  At first we thought it might be due to poor fixation, since the centers of tightly-wound rolls were affected, but we also observed this in the outer parts of loosely wound rolls.  I soak the blocks before sectioning; could non-uniform swelling result in variations of the section thickness?  (These are 7 microns thick.)
>
> Apologies if this information is available somewhere else; I tried unsuccessfully searching the archives.
>
> Thank you,
>
> Julie Cohen
>
> Research Lab Tech
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
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