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

Techniques for Imaging Ca2+ Signaling in Human Sperm

DOI:
10.3791/1996-v

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07:38 min

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June 16, 2010

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, , , , , , , ,
1School of Biosciences,University of Birmingham, 2School of Medicine,University of Birmingham, 3Centre for Human Reproductive Science,Birmingham Women’s Hospital

Chapters

  • 00:00Title
  • 00:50Introduction
  • 01:26Preparation and Staining of Sperm Cells
  • 04:32Imaging Sperm Intracellular Ca Signals
  • 06:20Single-cell Fluorescence Intensity Data and its Relationship to Free Calcium
  • 07:06Conclusion

Summary

Automatic Translation

Automatic Translation

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.

Tags

Fluorescence MicroscopyCa2+ SignalingHuman SpermFluorescent DyesLoading MethodPhysiological StimulationCapacitating ConditionsAM EsterCytoplasmVisible Wavelength DyesRatiometric RecordingAdvantages And DisadvantagesImaging ChamberPoly-D-lysine Coated CoverslipPerfusion ApparatusTime-lapse AcquisitionRegions Of InterestData Normalization

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