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Updated: Jun 9, 2025

Author Spotlight: Unraveling the Initial Activation of the Adaptive Immune Response for Therapeutic Intervention

Author Spotlight: Unraveling the Initial Activation of the Adaptive Immune Response for Therapeutic Intervention

Published on: October 4, 2024

Imaging Initial Ca2+ Microdomains in Primary T Cells.

Fynn Gerlach¹, Franziska Möckl¹, Dejan Kovacevic²

¹The Calcium Signalling Group, Department of Biochemistry and Molecular Cell Biology, University Medical Centre Hamburg-Eppendorf.

Journal of Visualized Experiments : Jove

|October 21, 2024

Summary

This summary is machine-generated.

Researchers developed a new imaging technique to detect rapid calcium (Ca2+) microdomains in T cells. This method visualizes fast cellular signals crucial for T cell activation and function.

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Last Updated: Jun 9, 2025

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Imaging Local Ca2+ Signals in Cultured Mammalian Cells

Imaging Local Ca2+ Signals in Cultured Mammalian Cells

Published on: March 3, 2015

Area of Science:

Immunology
Cell Biology
Biophysics

Background:

Local, sub-second calcium (Ca2+) signals, or Ca2+ microdomains, are critical for T cell activation and fate.
These dynamic signals are initiated by NAADP binding to specific proteins and receptors on intracellular Ca2+ stores.
Existing methods struggle to capture the high temporal and spatial resolution of these rapid Ca2+ events.

Purpose of the Study:

To develop and validate a high-resolution imaging technique for detecting rapid Ca2+ microdomains.
To enable the study of subcellular Ca2+ signaling dynamics in T cells.
To provide a tool for analyzing fast Ca2+ events in various cell types.

Main Methods:

Utilized a combination of two Ca2+ indicators, Fluo-4 AM and FuraRed AM, for high-resolution imaging.
Developed an open-source, semi-automated Ca2+ microdomain detection approach using Python.
Applied the technique to analyze fluorescence videos of primary murine and human T cells.

Main Results:

Successfully detected Ca2+ microdomains at the subcellular level in T cells with high temporal and spatial resolution.
The developed workflow reliably captures fast and dynamic Ca2+ signals.
Demonstrated the applicability of the method to other cell types, including NK cells and neuronal cell lines.

Conclusions:

The novel imaging and analysis workflow enables reliable detection of subcellular Ca2+ microdomains.
This technique provides unprecedented insight into the dynamics of Ca2+ signaling in T cell activation.
The method is versatile and applicable to a broader range of cell types for studying Ca2+ signaling.