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Updated: Dec 24, 2025
Simultaneous Measurement of Mitochondrial Calcium and Mitochondrial Membrane Potential in Live Cells by Fluorescent Microscopy
Published on: January 24, 2017
Mitochondria determine the sequential propagation of the calcium macrodomains revealed by the super-resolution
Yulin Zhang1, Jianyong Wang1, Shijia Xing1
1State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, Peking University, Beijing, 100871, China.
Abstract:
Despite the wide application of super-resolution (SR) microscopy in biological studies of cells, the technology is rarely used to monitor functional changes in live cells. By combining fast spinning disc-confocal structured illumination microscopy (SD-SIM) with loading of cytosolic fluorescent Ca2+ indicators, we have developed an SR method for visualization of regional Ca2+ dynamics and related cellular organelle morphology and dynamics, termed SR calcium lantern imaging. In COS-7 cells stimulated with ATP, we have identified various calcium macrodomains characterized by different types of Ca2+ release from endoplasmic reticulum (ER) stores. Finally, we demonstrated various roles of mitochondria in mediating calcium signals from different sources; while mitochondria can globally potentiate the Ca2+ entry associated with store release, mitochondria also locally control Ca2+ release from the neighboring ER stores and assist in their refilling processes.
Insights
Super-resolution microscopy now visualizes live cell calcium dynamics using SR calcium lantern imaging. This technique reveals how mitochondria interact with endoplasmic reticulum calcium stores during cellular signaling.
Area of Science:
- Cell Biology
- Microscopy
- Biophysics
Background:
- Super-resolution (SR) microscopy is vital for cellular studies but underutilized for live-cell functional monitoring.
- Visualizing dynamic intracellular processes like calcium signaling requires advanced imaging techniques.
Purpose of the Study:
- To develop a super-resolution method for observing regional calcium dynamics and organelle behavior in live cells.
- To investigate the interplay between calcium release, organelle morphology, and mitochondrial function.
Main Methods:
- Combined fast spinning disc-confocal structured illumination microscopy (SD-SIM) with fluorescent calcium indicators.
- Developed SR calcium lantern imaging for high-resolution visualization of live cell calcium transients.
- Utilized COS-7 cells stimulated with ATP to induce calcium signaling.
Main Results:
- Identified distinct calcium macrodomains characterized by varied calcium release patterns from endoplasmic reticulum (ER) stores.
- Demonstrated mitochondria's multifaceted roles in mediating calcium signals.
- Observed mitochondria globally potentiating calcium entry and locally controlling ER calcium release and refilling.
Conclusions:
- SR calcium lantern imaging enables unprecedented visualization of live-cell calcium dynamics and organelle interactions.
- Mitochondria play critical roles in modulating both global and local calcium signaling events within the cell.
- The study provides new insights into the spatial and temporal regulation of intracellular calcium homeostasis.
