Re: Some help for a first time user
Dave Coder (dave@nucleus.immunol.washington.edu)
Fri, 5 Nov 93 11:44:26 -0800
I spent about 6 weeks of 10 hour days developing a protocol for stripping colonic
epithelium from rat colon. This was fresh tissue; if fixed it's going to very hard
to do. The following should have the details of the method (it's been so long I
can't quite remember):
Author: Lupton-J-R. Coder-D-M. Jacobs-L-R.
Title: Influence of luminal pH on rat large bowel epithelial cell
cycle.
Source: Am-J-Physiol. 1985 Sep. 249(3 Pt 1). P G382-8.
Several tricks included: diothiothreitol to cut up the mucus, use of an EDTA soak to
loosen cell-cell junctions followed by vibration to dislodge epithelium, washing to
get rid of debris. Ethanol seemed to work for a fixative; PI worked quite well for
staining: good G0,1 CVs were 4%.
It's a shitty business, but somebody has to do it.
Dave Coder
Internet: dcoder@u.washington.edu
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