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Techniques for Imaging Ca2+ Signaling in Human Sperm

DOI :

10.3791/1996-v

7:38 min

June 16th, 2010

June 16th, 2010

16,863 Views

1School of Biosciences, University of Birmingham, 2School of Medicine, University of Birmingham, 3Centre for Human Reproductive Science, Birmingham Women’s Hospital

Stimulus-evoked [Ca2+]i signals of individual human sperm are assessed. Motile cells are loaded with Ca2+-sensitive fluorescent dye (AM-ester method) and immobilised in a perfusable chamber. Cells are imaged by time-lapse fluorescence microscopy and stimulated via the perfusing medium. Responses of single cells (or regions) are analysed offline using Excel.

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Fluorescence Microscopy

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