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Updated: May 15, 2026

Imaging Local Ca2+ Signals in Cultured Mammalian Cells

Imaging Local Ca²⁺ Signals in Cultured Mammalian Cells

Published on: March 3, 2015

Highlightable Ca2+ indicators for live cell imaging.

Hiofan Hoi¹, Tomoki Matsuda, Takeharu Nagai

¹Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada T6G 2G2.

Journal of the American Chemical Society

|December 22, 2012

Summary

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Researchers developed a novel fluorescent protein tool combining highlighter and calcium indicators. This allows spectral marking of single cells and imaging of their calcium dynamics, advancing cellular research.

Area of Science:

Biochemistry
Molecular Biology
Cell Biology

Background:

Fluorescent protein (FP) technology offers powerful tools for biological research, notably "highlighters" for spectral marking and calcium ion (Ca(2+)) indicators for dynamic imaging.
A significant advancement would be a single construct merging these functionalities, enabling highlightable Ca(2+) indicators for spectrally defined cellular marking and intracellular Ca(2+) dynamics imaging.

Purpose of the Study:

To create a novel hybrid fluorescent protein construct combining highlighter properties with Ca(2+) indication capabilities.
To develop a tool for spectrally marking individual cells within a tissue and simultaneously imaging their intracellular Ca(2+) dynamics.

Main Methods:

Exploration of three distinct protein design strategies for creating the hybrid FP.

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Dissection of Local Ca²⁺ Signals in Cultured Cells by Membrane-targeted Ca²⁺ Indicators

Published on: March 22, 2019

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Imaging Local Ca2+ Signals in Cultured Mammalian Cells

Imaging Local Ca²⁺ Signals in Cultured Mammalian Cells

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Successful strategy involved creating a circularly permuted green-to-red photoconvertible FP and integrating it into a G-CaMP-type Ca(2+) indicator.

Optimization of the construct through directed evolution.

Main Results:

Identification of two optimized variants with excellent photoconversion properties.
Demonstrated up to a 4.6-fold increase in red fluorescence intensity upon Ca(2+) binding.
Successful utility validation in HeLa cells and rat hippocampal neurons.

Conclusions:

The developed hybrid FP serves as a highlightable Ca(2+) indicator, enabling spectrally defined single-cell marking and Ca(2+) dynamics imaging.
These variants represent a significant advancement in fluorescent protein technology for cellular research.
The tool is effective in diverse cell types, including neurons, highlighting its broad applicability.