Recording and sorting live human sperm undergoing acrosome reaction
Fluorescent Pisum sativum agglutinin rapidly labels and stabilizes the acrosomal matrix when exocytic fusion pores open in live human sperm, as assessed by confocal microscopy, electron microscopy, and flow cytometry.
Felipe Carlos Martín Zoppino, Ph.D., Narciso D. Halón, B.S., Matías A. Bustos, M.S., Martín A. Pavarotti, Ph.D., Luis S. Mayorga, Ph.D.
Vol 97, Issue 6 , Pages 1309-1315
To develop and evaluate a method to detect acrosome reaction (AR) in live human sperm.
Basic research laboratory.
Human semen samples with normal parameters obtained from healthy donors.
Acrosome reaction assays.
Main Outome Measure(s):
Fluorescence assessment of AR.
Evaluating acrosomal exocytosis in live human sperm is challenging. In this study, we report that in reacting sperm, Pisum sativum agglutinin conjugated to fluorescein isothiocyanate rapidly permeates into the acrosome when fusion pores open and stabilizes the acrosomal matrix, preventing the dispersal of the granule contents.
Fluorescent Pisum sativum agglutinin can be used to visualize AR in real time, to determine the percentage of sperm undergoing exocytosis upon stimulation, and to separate the population of reacting sperm by flow cytometry.
Read the full text at: http://www.fertstert.org/article/S0015-0282(12)00289-0/fulltext