Techniques for Imaging Ca²⁺ Signaling in Human Sperm

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.