Mitochondrial Calcium Measurement

Mitochondrial calcium measurement is the quantitative assessment of Ca2+ levels or flux within mitochondria, a key approach for studying how cells regulate energy production, signaling, and stress responses. Researchers typically use genetically encoded or chemical fluorescent calcium indicators targeted to mitochondria; calcium binding changes the indicator’s fluorescence, allowing signals to be tracked as Ca2+ enters through channels such as the mitochondrial calcium uniporter and is released or buffered. These measurements reveal how calcium uptake interacts with the mitochondrial membrane potential, respiration, and cell survival, supporting research on metabolism, muscle and neuronal function, apoptosis, and mitochondrial disease.

Mitochondrial Calcium Measurement - Related Videos

Research

JoVE EoE - Assay Techniques

Calcium Influx Assay to Measure Mitochondrial Calcium Uptake in Cultured Cells

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2025

This video describes a confocal microscopy imaging-based assay to measure mitochondrial calcium uptake in cultured cells. The assay uses the calcium-sensitive fluorescent dye Rhod-2/AM to measure the mitochondrial calcium in permeabilized cultured cells.

Simultaneous Measurement of Mitochondrial Calcium and Mitochondrial Membrane Potential in Live Cells by Fluorescent Microscopy

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Cited by 29 •

2017

Mitochondria can utilize the electrochemical potential across their inner membrane (ΔΨm) to sequester calcium (Ca2+), allowing them to shape cytosolic Ca2+ signaling within the cell. We describe a method for simultaneously measuring mitochondria Ca2+ uptake and ΔΨm in live cells using fluorescent dyes and confocal microscopy.

Mitochondrial Calcium Uptake Assay: A Plate Reader-Based Method for Measuring Calcium Uptake in Isolated Mitochondria

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2025

This video describes a plate reader-based assay to measure mitochondrial calcium uptake in isolated mitochondria. The assay analyzes the fluorescence signal of a calcium-sensitive fluorescent dye to study the kinetics of mitochondrial calcium uptake and calcium overload.

Research

JoVE Journal - Biology
Free Sample

Monitoring Dynamic Changes In Mitochondrial Calcium Levels During Apoptosis Using A Genetically Encoded Calcium Sensor

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Cited by 10 •

2011

This protocol describes a method for real-time measurement of mitochondrial calcium fluxes by fluorescent imaging. The method takes advantage of a circularly permutated YFP-based dual-excitation ratiometric calcium sensor (ratiometric pericam-mt) selectively expressed in mitochondria.

Measuring Fast Calcium Fluxes in Cardiomyocytes

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Cited by 3 •

2011

We present a method to isolate rapid (microsecond) calcium events from slower fluxes in living cells using laser scanning confocal microscopy. The method measures fluorescence intensity fluctuations of calcium indicators by recording line scans of several hundred pixels in a cell. Histogram analysis allows us to isolate the time scales of different calcium fluxes.

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