easiest oxidative burst assay

Sarah Vowells (svowells@nih.gov)
Mon, 5 Aug 1996 09:59:40 -0400 (EDT)

>> I'm looking at phagocytosis and oxidative burst in porcine neutrophils.
>> I plan on performing a simultaneous assay using FITC-zymosan and
>> hydroethidium, and will be processing lots of samples by flow cytometry.
>> I would like to fix the cells in PFA after phagocytosis - will fixation
>> affect the resulting flourescence of EtBr produced during the oxidative
>> burst?

Something else you may want to consider for evaluation of the
oxidative burst is using the dihydrorhodamine assay. While I wouldn't
recommend using it concurrently with FITC zymosan, it is a fast, simple
assay (about an hour, from start to finish), and is very specific for
superoxide and/or hydrogen peroxide production.

Here is the reference...

Flow Cytometric Analysis of the Granulocyte Respiratory Burst: A
Comparison Study of Fluorescent Probes.

Journal of Immunological Methods 178 (1995): 89-97

Sarah Vowells
svowells@nih.gov
NIAID
National Institutes of Health

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