Myco and DAPI - revisited

David L. Haviland, Ph.D. dhavilan at IMM2.IMM.UTH.TMC.EDU
Fri Jan 10 15:35:18 EST 1997


Hi:

I'm a little confused as to why indicator cells such as 3T6 would be
employed in using DAPI for the dectection of mycoplasma.  I've had a paper
by Russel, WC, Newman, C,  & Williamson, DH. 1975.  "A simple cytochemical
technique for demonstration of DNA in cells infected with mycoplasmas and
viruses." Nature 253:461-461.

In this protocol, cells are cultures as they normally would, and if
possible, on glass.  Cells are then stained with DAPI and then observered
directly with a fluorescent microscope that has optics for 450 nm emission
(365 nm excitation).  Mycoplasma is readily seen in the cytoplasm and on
the surface of infected cells.  Uninfected cells only have signal in the
nucleus.

So my question is that an indicator cell will obviously work just fine, but
why not scan for mycoplasma directly on the cells in question?

For the record, I haven't had access to a fluoresent scope until now...
otherwise I have had reasonable detection of mycoplasma using the Stragene
PCR myco-tect system.  We used to use the BRL kit but it took to long.

David

=============================
 David L. Haviland, Ph.D.
 Asst. Prof. Immunology 
 University of Texas - Houston, H.S.C.
 Institute of Molecular Medicine  
 2121 W. Holcombe Blvd.  
 Houston, TX  77030 
 Internet:"dhavilan at imm2.imm.uth.tmc.edu" 
 Voice: 713.500.2413  FAX: 713.500.2424
 ------------------------------------------------------  
" Sometimes you're the windsheild, sometimes you're the bug."
=============================




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