Re: [Histonet] Re: Fluorescent DAB?????

From:Gayle Callis

I can't answer your question - take it up with John Kiernan 
per my email to Histonet.  This was used as a method to 
visualize myelinated nerves and fibers - John has more experience with the 
chemistry and use of this chromogen's fluorescence, and he did send at 
abstract to me for a European Neuroscience Association meeting in 2000.

The subject should certainly generate some interesting discussion though.

At 02:49 PM 6/26/2007, you wrote:
>Gayle Callis wrote: "DAB will fluoresce when excited at same wavelength as
>My questions are:  Assuming that all the parameters were kept  equivalent
>within a "batch" of slides:
>Can it be used in image analysis to quantify expression of a particular
>epitope of interest among different samples in a batch of stained slides?
>Or is DAB precipitate more of a random quantity depending on substrate
>availability/enzyme activity and therefore cannot be used to compare 
>among  slides
>(I realize that enzyme kinetics make the word "random" a bit of  an
>contradiction in this sense)?  This is based more upon the premise  that 
>the fluorescent
>secondary only has so many places to bind vs. HRP that  creates more color
>the longer it works......(at least until the reaction is  stopped or the
>substrate/cofactors are exhausted).
>Albert C.  Grobe, PhD
>International Heart Institute of Montana Foundation
>Tissue  Engineering Lab, Saint Patrick  Hospital

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610

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