Measuring DNA content

From:Gayle Callis

What do you mean by measuring? Quantitative or qualitative? 

Would Feulgen staining be more qualitative than other specific DNA stains
i.e. DAPI (UV excitation) or TOPRO3 (633 nm in far red) using image
analysis with confocal or other imaging methods? Feulgen does not require
any special fixation, although Bouins is usually avoided.  Methyl green is
also a DNA stain, as in methyl green pyronin, but Carnoys is preferred,
with NBF acceptable. Vinnie del Speranza wrote a wonderful publication in
Laboratory Medicine many years ago on the Feulgen reaction for DNA and HCl
hydrolysis of the molecule. It may be helpful to contact him.  I'm not sure
I have this on file anymore.     

Not familiar enough with our Zeiss 510 META CLSM at this time to tell you
more about analyzing quantitatively.  There is a confocal listserver with
people who do this type of thing with a high degree of image analysis
experience for fluorescence and confocal microscopy applications. Another
good source of information about this would be technical services from
Molecular Probes, they deal with many dyes that bind specifically to DNA
for fluorescence work. 

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
S. 19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367 (lab with voice mail)
406 994-4303 (FAX)


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