Journal
ANDROLOGIA
Volume 42, Issue 1, Pages 20-26Publisher
WILEY-BLACKWELL
DOI: 10.1111/j.1439-0272.2009.00949.x
Keywords
Acrosome reaction; 7-amino-actinomycin-D; flow cytometry; human spermatozoa; propidium iodide
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Funding
- National Research Foundation (NRF), Pretoria, South Africa [2054206]
- South African Bureau of Standards (SABS)
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P>Evaluation of the acrosome reaction can shed light on the fertilising competence of spermatozoa. To eliminate false-positive results when evaluating the acrosome status of human sperm cells, two viability probes propidium iodide (PI) and 7-amino-actinomycin D (7-AAD) were compared for their ability to stain nonviable cells post-fixation and permeabilisation. Both the mean fluorescence and % dead cells differed significantly with time (P < 0.0001). Unlike PI, 7-AAD did not leach from cells and fluorescence remained stable for up to 4 h. Furthermore, 7-AAD proved to be a proficient marker to exclude dead sperm cells during flow cytometric evaluation of ionophore-induced acrosome reaction.
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