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...

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Bibliographic Details
Published in:Journal of biotechnology 2008, Vol.133 (1), p.90-95
Main Authors: Micera, Elisabetta, Albrizio, Maria, Surdo, Nicoletta Concetta, Zarrilli, Antonia
Format: Article
Language:English
Subjects:
agar
Agar matrix
analytical methods
Animals
Biological and medical sciences
Biotechnology
Calcium
Calcium - metabolism
Cell Separation - methods
Cell Survival
Cells, Cultured
Cells, Immobilized - physiology
cytochemistry
cytoplasm
Equine
Flow Cytometry - methods
Fundamental and applied biological sciences. Psychology
Horses
image analysis
Image Enhancement - methods
Image Interpretation, Computer-Assisted - methods
intracytoplasmic calcium
Male
Microscopy, Fluorescence - methods
Microscopy, Video - methods
microspectrofluorimetry
Semi-immobilization
Spermatozoa
Spermatozoa - cytology
Spermatozoa - metabolism
videoimaging
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Summary: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.
ISSN:0168-1656
1873-4863
DOI:10.1016/j.jbiotec.2007.09.011