Fixatives

Julia Parker lancekas at u.washington.edu
Mon Apr 24 17:28:12 EST 1995


I am currently trying to differentiate between intracellular and 
extracellularly-localized bacteria.  I am studying obligate intracellular 
parasites, thus I cannot use the gentamicin protection assay.  I was 
trying to use a 1% paraformaldehyde fixation to crosslink the membrane 
and label extracellular organisms and then come back with detergent to  
introduce holes before staining the intracellular bugs.  I have been 
using epi-fluorescence to visualize and photograph the process.  Recently 
I have been told that the fixation itself is enough to knock large enough 
holes in the membrane to allow antibody to stain intracellular organisms 
as well as extracellular ones before I ever apply detergent to 
solubilize.  Can anyone with expertise in the area, give me advise about a 
better technique or modification to fit my purpose?  I would appreciate your 
advise very much.

Sincerely, Julia

Please e-mail me directly at lancekas at u.washington.edu



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