On 12/14/2011 9:55 AM, Elizabeth Cameron wrote:
We are doing a Hale's colloidal iron stain (no counterstain) on serial sections
of mouse kidneys. We are staining an entire kidney at a time (about 90-150
slides), and after many successful runs, we are now finding some slides in each
batch with very uneven staining. Half of a section will stain as it should,
and the rest of the section is very pale. It seems to be in a similar area
from one section to the next, but not exactly the same area. It does not look
like a deparaffinization issue. There may be two or three slides in a row like
this, or just one section on a slide, followed by 30-40 that look fine. The
sections are 6 microns. Any ideas on why this might be happening? Thanks!
Elizabeth M. Cameron, HT, QIHC (ASCP)
The Jackson Laboratory
Bar Harbor, Maine
(207) 288-6561
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Hi Elizabeth:
It seems like every time someone posts an "uneven staining" question,
the solution (not just the answer) is always incomplete deparafinization.
Try fresh xylene, minimum 2 changes 5 minutes each, and fewer slides in
the rack and some agitation (of the slides, not of you).
If that does not solve the problem incomplete fixation might be the
culprit.Perfused tissue? Fixed how long in ?
Geoff
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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
mcaul...@umdnj.edu
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