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Related Experiment Video

Updated: Jun 12, 2026

Techniques for Imaging Ca2+ Signaling in Human Sperm

Techniques for Imaging Ca²⁺ Signaling in Human Sperm

Published on: June 16, 2010

Techniques for imaging Ca2+ signaling in human sperm.

Katherine Nash¹, Linda Lefievre, Ruben Peralta-Arias

¹School of Biosciences, University of Birmingham, USA.

Journal of Visualized Experiments : Jove

|June 23, 2010

Summary

This summary is machine-generated.

This study details a method for measuring calcium (Ca2+) signaling in human sperm using fluorescent dyes. The technique involves dye loading, imaging, and data analysis for observing Ca2+ dynamics during physiological stimulation.

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Medium-throughput Screening Assays for Assessment of Effects on Ca2+-Signaling and Acrosome Reaction in Human Sperm

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Last Updated: Jun 12, 2026

Techniques for Imaging Ca2+ Signaling in Human Sperm

Techniques for Imaging Ca²⁺ Signaling in Human Sperm

Published on: June 16, 2010

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

Published on: May 24, 2013

Medium-throughput Screening Assays for Assessment of Effects on Ca2+-Signaling and Acrosome Reaction in Human Sperm

Medium-throughput Screening Assays for Assessment of Effects on Ca²⁺-Signaling and Acrosome Reaction in Human Sperm

Published on: March 1, 2019

Area of Science:

Cell biology
Biochemistry
Reproductive science

Background:

Calcium (Ca2+) signaling is crucial for cellular functions, including sperm motility and fertilization.
Fluorescence microscopy with Ca2+-sensitive dyes is a powerful tool for studying Ca2+ dynamics in live cells.

Purpose of the Study:

To describe a reliable method for loading human sperm with Ca2+-reporting dyes.
To establish a protocol for measuring Ca2+ signaling in human sperm during physiological stimulation.

Main Methods:

Human sperm are isolated and incubated under capacitating conditions.
Sperm are loaded with cell-permeant acetoxy methyl ester (AM) Ca2+-reporting dyes.
Time-lapse fluorescence microscopy is used to record Ca2+ signals, followed by offline data analysis.

Main Results:

A detailed protocol for Ca2+ dye loading and imaging in human sperm is presented.
The method allows for the measurement of spatial and temporal Ca2+ changes in individual sperm cells.
Data normalization provides Ca2+ responses as a percentage change in fluorescence.

Conclusions:

This fluorescence microscopy method provides a robust approach for studying Ca2+ signaling in human sperm.
The described technique facilitates the investigation of Ca2+ dynamics underlying sperm function.
Understanding Ca2+ signaling in sperm is vital for reproductive biology research.