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Single-Cell Calcium Imaging for Studying the Activation of Calcium Ion Channels

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A fluorescent activatable probe for imaging intracellular Mg²⁺ .

Ryan Treadwell¹, Fabio de Moliner, Ramon Subiros-Funosas

¹Medical Research Council Centre for Inflammation Research, The University of Edinburgh, EH16 4TJ Edinburgh, UK. marc.vendrell@ed.ac.uk.

Organic & Biomolecular Chemistry

|December 20, 2017

Summary

A new BODIPY probe enables sensitive detection and imaging of free magnesium ions (Mg²⁺) in cells. This probe distinguishes Mg²⁺ from calcium ions (Ca²⁺), offering enhanced fluorescence for accurate biological studies.

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Area of Science:

Chemical Biology
Molecular Imaging
Biochemistry

Background:

Accurate detection of free magnesium ions (Mg²⁺) is crucial for understanding cellular processes.
Existing detection methods can suffer from interference by other biologically relevant ions, such as calcium (Ca²⁺).
Development of selective and sensitive probes for Mg²⁺ is an ongoing challenge in chemical biology.

Purpose of the Study:

To develop and characterize a novel activatable BODIPY probe for the selective detection and fluorescence imaging of free Mg²⁺.
To demonstrate the probe's ability to function without interference from Ca²⁺.
To investigate the mechanism behind the probe's fluorescence amplification upon Mg²⁺ binding.

Main Methods:

Synthesis and characterization of a novel BODIPY-based fluorescent probe incorporating a quinolizine-based chelator.
In vitro fluorescence spectroscopy to assess probe selectivity and sensitivity towards Mg²⁺ in the presence of Ca²⁺.
Confocal fluorescence microscopy for cell imaging studies to visualize intracellular Mg²⁺ distribution.

Main Results:

The developed BODIPY probe exhibits high selectivity and sensitivity for free Mg²⁺, with minimal response to Ca²⁺.
Upon binding to physiological concentrations of Mg²⁺, the probe shows significant fluorescence amplification.
The fluorescence enhancement is attributed to the restricted intramolecular rotation of the fluorophore upon chelation by the quinolizine core.
Successful in vitro cell imaging of Mg²⁺ was achieved, demonstrating the probe's utility in biological systems.

Conclusions:

A novel activatable BODIPY probe has been successfully developed for selective and sensitive detection of free Mg²⁺.
The probe's design effectively prevents interference from Ca²⁺, enhancing its reliability for biological applications.
This probe represents a valuable tool for in vitro fluorescence cell imaging and studying the biological roles of Mg²⁺.