A semi-immobilizing system associated with microspectrofluorimetric and videoimaging analysis for intracytoplasmic calcium measurement in individual viable spermatozoa.
We describe a fast and simple method to monitor variations of intracytoplasmic calcium concentrations in very small and motile cells such as mammalian spermatozoa during measurement time. The method combines a procedure of sperm cells semi-immobilization with microspectrofluorimetric measurement of intracytoplasmic calcium concentrations supported by videoimaging that allows also a constant monitoring of viability during the time of calcium recording. In this paper we show that a semi-immobilization of individual viable spermatozoa can be obtained preparing the agar matrix in a two step procedure. The cell immobilizing system proposed, associated with the microspectrofluorimetric analysis supported by videoimaging, is a simple, rapid and useful tool for those studies having the goal of correlating the presence and cellular distribution of ion channels with their functional status and their response to physiologic and/or pharmacological molecules.
Micera
Elisabetta
E
Department of Animal Production, Faculty of Veterinary Medicine, University of Bari, strada prov.le per Casamassima km 3, 70010 Valenzano, Bari, Italy.
Albrizio
Maria
M
Surdo
Nicoletta Concetta
NC
Zarrilli
Antonia
A
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival; D002478:Cells, Cultured; D018914:Cells, Immobilized / Q000502:physiology; D005434:Flow Cytometry / Q000379:methods*; D006736:Horses; D007089:Image Enhancement / Q000379:methods; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D008297:Male; D008856:Microscopy, Fluorescence / Q000379:methods*; D018715:Microscopy, Video / Q000379:methods*; D013094:Spermatozoa / Q000166:cytology* / Q000378:metabolism*
D002118:Calcium