FW: Mitotic index - McAuliffe cont'd
|From:||"Monson, Frederick C." |
> From: Monson, Frederick C.
> Sent: Tuesday, June 4, 2002 8:54 AM
> To: 'McAuliffe, Geoff'
> Cc: 'List-Microscopy'
> Subject: RE: Mitotic index - McAuliffe cont'd
> I agree with Geoff, Augustin. Br-UdR is the current way to go. It
> actually requires more work than 3H-TdR, but for short-term experiments,
> its use avoids a LOT of safety and regulatory hassles. Problem is that in
> longer term experiments, where 3H-TdR is known, by literature support, to
> 'breed true', there appears to be less such support for long-term Br-UdR.
> If you just count the layers (i.e., the number of steps to finished
> product), 3H-TdR comes out way ahead in resolution, ease of use and
> precision, BUT the hassle in use of radioactive labels is almost totally
> limiting if one only considers the added cost of handling the remains.
> When I used 3H-TdR on rabbit urinary bladder in situ(vivo) in the early
> 90's, I ended up using very low dosage per gram of body weight (~0.1uCi/g)
> when I introduced the label i.v. and normal (for me!) dosage (0.5uCi/ml or
> g) in aerated Hank's BSS (NO BSA added!!) for in vitro incubations of
> either whole bladder or bladder strips. Labeling indices proved to be
> similar in three categories: whole bladder labeled either in vivo(1) or
> in vitro(2) and bladder strips in vitro.
> NOTE: the throw-away radiation level was 0.05uCi/g body weight at that
> time (early '90's) so even when dosed less and exposed in a special
> manner, hazardous waste disposal was still required for the carcass.
> Needless to say, the growing cost of disposal caused me to expend both
> energy and time in developing a means by which I could dose with
> 0.049uCi/g body weight and not have to wait for a generation (mine!) for
> the autoradiograms to expose. Got close, but time and money ran out.
> Fred Monson
> Frederick C. Monson, PhD
> Center for Advanced Scientific Imaging
> Schmucker II Science Center
> West Chester University
> South Church Street and Rosedale
> West Chester, Pennsylvania, USA, 19383
> Phone: 610-738-0437
> FAX: 610-738-0437
> CASI URL: http://darwin.wcupa.edu/casi/
> WCUPA URL: http://www.wcupa.edu/
> Visitors URL: http://www.wcupa.edu/_visitors/
> From: Geoff McAuliffe
> Sent: Monday, June 3, 2002 4:58 PM
> To: Agustín Venzano
> Cc: HistoNet Server
> Subject: Re: Mitotic index
> I believe that bromodeoxyuridine is a more accurate index of mitotic
> activity, but I can't remember the citation right now. Actually seeing
> figures is difficult, they don't last long! S-phase often lasts 10-12
> mitosis lasts 20 minutes so incorporation of something into the DNA during
> S-phase is the way to go. The "old" way with tritiated thymidine
> has problems due to disposal of animals and contaminated reagents.
> Agustín Venzano wrote:
> > Dear netters: I'm planning a sampling of ruminal papillae (i.e. special
> > structures endowed with epithelium and lamina propria located in the
> > forestomach of ruminants) in young cattle. The project is aimed at
> > the growth rate of these structures specialised in nutrients absorption,
> > it is necessary to estimate the mitotic index. My questions are:
> > 1.What staining would you prefer for seeing DNA and mitosis?
> > 2. Do you consider c-kit to be an adequate marker of the mitotic rate
> > through IHC in paraffin blocks?
> > Thank you in advance
> > Sincerely yours
> > Agustin Jose Venzano Halliburton
> > DVM-Pathology Group
> > INTA, Argentina
> Geoff McAuliffe, Ph.D.
> Neuroscience and Cell Biology
> Robert Wood Johnson Medical School
> 675 Hoes Lane, Piscataway, NJ 08854
> voice: (732)-235-4583; fax: -4029
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