> ----------
> 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.
> 
> Regards,
> 
> 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
> fmonson@wcupa.edu
> 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
> 
> Augustin:
> 
>     I believe that bromodeoxyuridine is a more accurate index of mitotic
> activity, but I can't remember the citation right now. Actually seeing
> mitotic
> figures is difficult, they don't last long! S-phase often lasts 10-12
> hours,
> 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
> autoradiography
> 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
> largest
> > forestomach of ruminants) in young cattle. The project is aimed at
> defining
> > the growth rate of these structures specialised in nutrients absorption,
> so
> > 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
> --
> **********************************************
> 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
> mcauliff@umdnj.edu
> **********************************************
> 
> 
> 
> 
> 
>