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
EM Core Facility
Weill Cornell Medical College
1300 York Avenue, Room A-105
New York, NY 10021
lab: 212-746-6146
email: juc2...@med.cornell.edu<mailto:juc2...@med.cornell.edu>
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732) 235-4583; fax: -4029
mcaul...@rwjms.rutgers.edu
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