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Fig. 6. Passage through inert polycarbonate filters induces the sperm acrosome
reaction. (A) Capacitated Acr3-EGFP sperm
(2.1x104) were placed in the lower chamber and incubated for
30 minutes. The sperm acrosome status was assayed before (lower chamber) and
after (upper chamber) penetration of individual polycarbonate filters (pore
size, 1.2-12 µm); two example filters are imaged on the right. (B)
Fixed sperm were stained with propidium iodide and divided into
acrosome-intact and -reacted populations by FACS (left). The acrosome status
of each population was confirmed by confocal microscopy (right). (C)
Acr3-EGFP sperm, isolated before (lower chamber) and after (upper
chamber) passage through individual filters (pore size, 1.2-12 µm), were
fixed, stained with propidium iodide and scored by FACS for acrosome status.
(D) Epididymal sperm were isolated and capacitated for 1 hour and an
aliquot was passed over a glass bead column to remove acrosome-reacted sperm.
The acrosome status of epididymal (left) and glass-bead-treated (right) sperm
was assayed by FACS before (lower chamber) and after (upper chamber) passage
through a 3 µm pore-size polycarbonate filter.
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