Re: EMA Staining Protocol
(no name) ((no email))
Fri, 11 Nov 1994 08:18:46
Mark Rehse writes:
> I have been working with 5 uM ethidium monoazide (EMA) trying to identify
> nonviable cells in a 1% PFA fixed population of cells stained with fitc
> and pe conjugated antibodies. Problems I've had are dramatic changes in
> light scatter due to either the EMA or the DMSO used to solubilize the
> EMA, as well as a gross underestimate of nonviability in comparison to
> the same cells unfixed and PI stained. Additionally, the EMA induces a
> high fluorescent background. Anyone have any suggestions as to what stock
> concentration to make the EMA up in and how to solubilize it? Do people
> out there use EMA for this application of is there a better stain? Thanks
> in advance.
> Mark Rehse Rehsema@cellpro.cellpro.com
Carl Stewart has a protocol in the Handbook of Flow Cytometry Methods
page 174 which uses 5ug/ml EMA.
Paul
J.Paul Robinson, Purdue University Cytometry Labs
CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories
and distributed free of charge as an educational service to the cytometry community.
If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL,
Purdue University, West Lafayette, IN 47907. Phone:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu
EMAIL robinson@flowcyt.cyto.purdue.edu
