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{I am trying to determine the cycling population of CD8 T cells in vivo. As
you know, BrdU labelling involves the fixation of cells for staining, and I
am wondering if anyone out there knows any alternative labelling methods
which do not require the fixation of cells. }
A simple method is to use BrdU, but use the BrdU queching of Hoechst 33342 in
conjunction with mithramycin labeling to infer the amount of BrdU
incorporated. The most elegant way to analyze the data is to subtract the
Hoechst signal from the mithramycin signal for each cell and plotting the
difference vs. the mithramycin signal. See the illustrations in Tobey, Oishi,
and Crissman (J. Cell Physiol. 139:432-440).
Requires: dual laser system (457nm and uv excitation)
{(wondering when spring comes here)
the cherry trees by Portage Bay are now in blossm here.
Dave Coder
dcoder@u.washington.edu
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