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Techniques for Imaging Ca2+ Signaling in Human Sperm
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Techniques for Imaging Ca2+ Signaling in Human Sperm

Article DOI: 10.3791/1996-v • 07:39 min • June 16th, 2010
June 16th, 2010 •
1, 2, 1, 1, 1, 2, 1, 3, 1
1School of Biosciences, University of Birmingham, 2School of Medicine, University of Birmingham, 3Centre for Human Reproductive Science, Birmingham Women’s Hospital

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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 Ca2+ Signaling Human Sperm Fluorescent Dyes Loading Method Physiological Stimulation Capacitating Conditions AM Ester Cytoplasm Visible Wavelength Dyes Ratiometric Recording Advantages And Disadvantages Imaging Chamber Poly-D-lysine Coated Coverslip Perfusion Apparatus Time-lapse Acquisition Regions Of Interest Data Normalization
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