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Updated: Jul 3, 2026
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Ratiometric Calcium Imaging of Individual Neurons in Behaving Caenorhabditis Elegans
Published on: February 7, 2018
Electron-multiplying charge-coupled detector-based bioluminescence recording of single-cell Ca2+
Kelly L Rogers1, Jean-Rene Martin, Olivier Renaud
1Plate-forme d'Imagerie Dynamique, Imagopole,Institut Pasteur, Paris, France. krogers@pasteur.fr
Journal of Biomedical Optics
|July 8, 2008
Summary
New imaging technology enables long-term monitoring of intracellular calcium concentration ([Ca2+]i) in single cells. This breakthrough overcomes limitations of previous methods, facilitating easier and more stable bioluminescence resonance energy transfer (BRET) experiments.
Area of Science:
- Biochemistry
- Cell Biology
- Biophysics
Background:
- Genetically encoded intracellular calcium concentration ([Ca2+]i) indicators have faced challenges in translating initial excitement into scientific advances.
- Recombinant apo-aequorin from Aequorea victoria was the first genetically targeted Ca2+ sensitive protein.
- Bioluminescence resonance energy transfer (BRET) between aequorin and fluorescent proteins (like GFP) offers improved light intensity and stability.
Purpose of the Study:
- To report the use of electron-multiplying charge-coupled-detector (EMCCD) imaging for BRET measurement of single-cell [Ca2+]i.
- To demonstrate facile long-term monitoring of [Ca2+]i at the single-cell level using novel technology.
Main Methods:
- Utilized electron-multiplying charge-coupled-detector (EMCCD) imaging camera technology.
- Mounted EMCCD cameras on bioluminescence or conventional microscopes.
- Employed BRET reporters constructed from aequorin and fluorescent protein variants.
Main Results:
- Successfully demonstrated BRET measurement of single-cell [Ca2+]i.
- Showcased facile long-term monitoring capabilities of [Ca2+]i at the single-cell level.
- Validated EMCCD technology as a viable alternative to image-photon-detectors (IPD) for dynamic BRET experiments.
Conclusions:
- EMCCD imaging technology significantly simplifies long-term, single-cell [Ca2+]i monitoring via BRET.
- This advancement obviates the need for expensive and complex IPD equipment.
- Facilitates broader application of BRET for studying intracellular calcium dynamics.
