[Histonet] inconsistent H&E staining

[Julie Cohen]

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
EM Core Facility
Weill Cornell Medical College
1300 York Avenue, Room A-105
New York, NY 10021
lab: 212-746-6146
email: juc2023 <@t> med.cornell.edu<mailto:juc2023 <@t> med.cornell.edu>