Flow cytometric evaluation of the acrosome reaction of human spermatozoa: a new method using a photoactivated supravitaI stain

Abstract

A flow cytometric assay using a double-stain method for the measurement of the acrosome reaction of human spermatozoa is described. The use of a stable photoactivated stain, ethidium monoazide, allowed evaluation of the viability of spermatozoa. This stain was more stable in fixed samples than propidium iodide, which is not bound covalently to DNA and is therefore removed readily during the washing procedure. The permeabilized acrosome was labelled with Pisum sativum agglutinin conjugated with fluoroisothiocyanate. Since this lectin binds to the acrosome and acrosomal contents, a decrease in the fluorescence intensity allows the cytometric evaluation of the acrosome reaction. Microscopic analysis and flow cytometric analysis were well correlated and cell sorting was performed to ensure the homogeneity of each different subpopulation encountered.

Damage assessment of the equine sperm membranes by fluorimetric technique

“…The possibility of association of fluorescent probes for the evaluation of spermatozoa has been described in the literature (Graham et al, 1990;Centola et al, 1990;Henley et al, 1994;Sukardi et al, 1997;Papaioannou et al, 1997;Arruda et al, 2002;Huo et al, 2002;Arruda et al, 2003;Nagy et al, 2003;Love et al, 2003;Celeghini et al, 2008). However, most of the techniques allow evaluating simultaneous only plasma and acrosomal membranes.…”

Purification of Lysophospholipase of Human Spermatozoa and its Implication in the Acrosome Reaction1

“…Lysophospholipase activity was measured after a 4-h incubation period. [24]. This technique uses flow cytometry to evaluate the acrosome reaction only on viable sperm, thus providing a highly accurate measurement of the acrosome reaction.…”

Assessment of bovine spermatozoa viability using different cooling protocols prior to cryopreservation

“…The combination of different fluorescent probes, as used in our work for simultaneous evaluation of changes in the spermatozoan plasma and acrosomal membranes and mitochondrial potential, has been reported previously (Henley et al, 1994;Huo et al, 2002;Nagy et al, 2003). Percentages of spermatozoan plasma membrane, acrosomal membrane and mitochondrial potential integrity were lower in the RD cooling curve ( The effect of cooling rate on spermatozoan [Ital J Anim Sci vol.9:e88, 2010]…”