[Histonet] mycoplasma staining

[Alicia Henn Ph.D.]

Hi All,
    I use a bisbenzimide stain for routine mycoplasma detection of cell 
cultures.  Lately, I've had problems with a bright blue haze covering 
dimly-stained cells. I've tried changing every step of the stain, but still 
get blue fog. I fix with methanol/acetone (I've tried formalin as well) 
stain the cells in chamberslides or in solution (for non-adherent cells) for 
30 minutes, wash, air-dry chamberslides, mount with anti-fade mounting 
medium (I've tried several). I can visualize fluorescent beads with 
exquisite detail, so my microscope is OK. Anyone had any experience with 
this? I don't want to have to go back to PCR-based detection as that was a 
nightmare.

Alicia Henn, Ph.D.
Vaccinex, Inc.