Re: Circulation time of BRDU in a rat
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From: | rschoonh@sph.unc.edu |
To: | JErickson@genetics.com (Jamie Erickson), HistoNet@Pathology.swmed.edu |
Reply-To: | |
Date: | Thu, 24 Jun 1999 09:46:43 -0400 (Eastern Daylight Time) |
Content-Type: | TEXT/PLAIN; CHARSET=US-ASCII |
Jamie,
We use the following:
Dissolve 20mg BrdU per 1 ml phosphate buffered sterile saline (pH 7.6), held at room temp. and
shielded from light. This is administered IP at 5 ml/1Kg. The animals are sac.'ed 1 hour after
injection. So much for the easy part.
As regards measuring proliferation labeling index (LI). There are several ways that you could
generate the results, number of labeled cells/ mm (or other length unit) or number pos. cells
divided by total cells counted per crypt. we have used both of these methods (still do). see :
The Effects of Exposure Route on DNA Adduct Formation and Cellular Proliferation by
1,2,3,-Trichloropropane;
D.La, R. Schoonhoven, N. Ito, J.A. Swenberg;
Toxicology and Applied Pharmacology 140, 108-114 (1996)
Most of the details (on how we measured the LI) are given in that article. Our IHC methods have
changed since then (darn that thar progress thing ;-)) By measuring migration I would set up a
time study over several days, no more than 7 as at that point ALL cells would be labeled. Do an IP
injection each day ie: day 1 sac day 1 , day 2 (rec'd 2 IP injections) sac, etc...
Measure either length of labeled cells along basement membrane from base of crypt on up or, num. of
cells per measurement unit.
Please feel free to call if I can be of any assistance.
-- Begin original message --
>
> HI All,
> I know this was discussed sometime ago but could someone tell me the circulation time for
> BRDU, I believe the dose of BRDU given was 500 ug/kg. I'm doing a study to evaluate BRDU stained
> slides of Rat colon and need this info. Also how do people evaluate these slides, is it by number
> of positive cells/ crypt (how many do you count /slides?) . There seems to be two things to
> evaluate in these slides migration up the Crypt and proliferation, what's the best way to
> evaluate both of these??
>
> If someone could help I'd really appreciate it.
>
> Jamie Erickson
>
-- End original message --
best regards,
Bob
Robert Schoonhoven
Laboratory of Molecular Carcinogenesis and Mutagenesis
Dept. of Environmental Sciences and Engineering
University of North Carolina
CB#7400
Chapel Hill, NC 27599
Phone
office 919-966-6343
Lab 919-966-6140
Fax 919-966-6123
Don't go around saying the world owes you a living; the world owes you
nothing; it was here first.
Mark Twain [Samuel Langhornne Clemens] (1835-1910)
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