Re: Alternative to BrdU labelling in vivo ?

David L. Haviland, Ph.D. (HAVILAND@kids.wustl.edu)
Wed, 08 Mar 1995 12:50:24 -0600 (CST)

>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. Thanks in advance !!

Aki:

Way back in Grad School, I was using an Epics V and was also trying to
examine cell cycling in T cells. I tried using the Hoechst (sp?) 33342 (also
maybe the 33258, but I really do think it was the 33342 - even after 9
years!). Needless to say, I was unable to get a classic growing cell cycle
histogram with Hoechst stains. THE only thing I did accomplish was being
able to roll the Epics in UV mode in about 20 minutes - that was all.

To complete the project, we wound up using Propidium Iodide via a fixation,
triton, RNAse like protocol. That gave us the picture purfect curve we
wanted to see...

Hope this helps,
David

>Aki Hoji (wondering when spring comes here)
>University of Wisconsin, Madison.

*wishing* for big snow but dissapointed at yet another wimpy mid-west
winter here in STL!


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