Comparison between propidium iodide and 7-amino-actinomycin-D for viability assessment during flow cytometric analyses of the human sperm acrosome

Date
2010, 2010
Authors
Falzone N.
Huyser C.
Franken D.R.
Falzone N.
Huyser C.
Franken D.R.
Journal Title
Journal ISSN
Volume Title
Publisher
Abstract
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. © 2009 Blackwell Publishing Ltd.
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. © 2009 Blackwell Publishing Ltd.
Description
Keywords
ionophore; propidium iodide; acrosome; acrosome reaction; adult; article; cell membrane permeability; clinical assessment; comparative study; controlled study; flow cytometry; fluorescence; human; human cell; male; staining; time; Acrosome; Acrosome Reaction; Adult; Dactinomycin; False Positive Reactions; Flow Cytometry; Fluorescent Dyes; Humans; Male; Microscopy, Fluorescence; Propidium; Spermatozoa, ionophore, propidium iodide, acrosome, acrosome reaction, adult, article, cell membrane permeability, clinical assessment, comparative study, controlled study, flow cytometry, fluorescence, human, human cell, male, staining, time, Acrosome, Acrosome Reaction, Adult, Dactinomycin, False Positive Reactions, Flow Cytometry, Fluorescent Dyes, Humans, Male, Microscopy, Fluorescence, Propidium, Spermatozoa
Citation
Andrologia
42
1
Andrologia
42
1