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Measuring Intracellular Ca2+ Changes in Human Sperm using Four Techniques: Conventional Fluorometry, Stopped Flow Fluorometry, Flow Cytometry and Single Cell Imaging
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Biologia
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JoVE Journal Biologia
Measuring Intracellular Ca2+ Changes in Human Sperm using Four Techniques: Conventional Fluorometry, Stopped Flow Fluorometry, Flow Cytometry and Single Cell Imaging

Measuring Intracellular Ca2+ Changes in Human Sperm using Four Techniques: Conventional Fluorometry, Stopped Flow Fluorometry, Flow Cytometry and Single Cell Imaging

DOI:
10.3791/50344-v

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19:26 min

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May 24, 2013

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, , , , , , ,
1Departamento de Genética del Desarrollo y Fisiología Molecular,Instituto de Biotecnología-Universidad Nacional Autónoma de México, 2Math and Sciences Department,Edison State College

Capitoli

  • 00:05Titolo
  • 02:38Sperm Sample Preparation by the Swim-up Method
  • 04:52Fluorescent Dye Loading for Ca2+ Measurements
  • 05:41Conventional Fluorometry
  • 06:59Stopped Flow Fluorometry
  • 08:30Flow Cytometry
  • 09:40Single Cell Imaging
  • 12:11Results: Progesterone Induces Intracellular Ca2+ Changes in Human Sperm Detected by Fluorometric Techniques
  • 17:59Conclusion

Summary

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Intracellular Ca2+ dynamics are very important in sperm physiology and Ca2+-sensitive fluorescent dyes constitute a versatile tool to study them. Population experiments (fluorometry and stopped flow fluorometry) and single cell experiments (flow cytometry and single cell imaging) are used to track spatio-temporal [Ca2+] changes in human sperm cells.

Tags

Ca2+ SignalingSpermFluorescent DyesFluorometryFlow CytometrySingle Cell ImagingCalcium DynamicsSperm FunctionCapacitationAcrosome ReactionSperm Motility

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