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Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors

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Quantitative Model for Reversibly Photoswitchable Sensors.

Beatrice Adelizzi¹, Vincent Gielen², Thomas Le Saux¹

¹PASTEUR, Département de Chimie, École Normale Supérieure, PSL University, Sorbonne Université, CNRS, 24, rue Lhomond, Paris 75005, France.

|February 10, 2021

Summary

This study introduces a kinetic model for reversibly photoswitchable sensors (rs-sensors) to improve quantitative analyte detection. The model helps analyze key parameters for robust sensor design and application, enhancing performance in sensing technologies.

Keywords:

biosensors calcium sensor kinetic model reversibly photoswitchable proteins reversibly photoswitchable sensors

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Area of Science:

Biochemistry
Chemical Engineering
Molecular Biology

Background:

Reversibly photoswitchable sensors (rs-sensors) offer a novel approach for quantitative analyte detection.
The complex kinetic response of rs-sensors to illumination necessitates careful design and application strategies.

Purpose of the Study:

To develop and validate a generic kinetic model for rs-sensors.
To identify and analyze key thermokinetic parameters influencing rs-sensor performance.
To establish a reliable methodology for parameter evaluation in rs-sensor applications.

Main Methods:

Exploitation of a generic kinetic model for rs-sensors.
Analysis of thermokinetic parameters including dissociation constants, kinetic rates, and photoswitching cross-sections.
Experimental validation using a novel calcium sensor.

Main Results:

The kinetic model successfully identifies critical thermokinetic parameters for rs-sensors.
A methodology for robust parameter evaluation and reliable use has been introduced.
Experimental testing confirmed the model's applicability on a calcium-sensing rs-sensor.

Conclusions:

The developed kinetic model provides a framework for understanding and optimizing rs-sensor behavior.
The proposed methodology enhances the reliability of parameter analysis for rs-sensor development.
This work facilitates the rational design and application of advanced photoswitchable sensing systems.