Techniques for Imaging Ca2+ Signaling in Human SpermKatherine Nash 1, Linda Lefievre 2, Ruben Peralta-Arias 1, Jennifer Morris 1, Aduen Morales-Garcia 1, Tom Connolly 2, Sarah Costello 1, Jackson C. Kirkman-Brown 3, Stephen J. Publicover 1
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.
