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Updated: Jul 10, 2025

Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation

Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation

Published on: February 8, 2015

Fluorescence to measure light intensity.

Aliénor Lahlou^1,2, Hessam Sepasi Tehrani³, Ian Coghill³

¹PASTEUR, Department of Chemistry, École Normale Supérieure, PSL University, Sorbonne University, CNRS, Paris, France. Alienor.Lahlou@sony.com.

|November 23, 2023

Summary

This summary is machine-generated.

Researchers developed new methods using dyes and fluorescent proteins to measure light intensity and spatial distribution in fluorescence microscopy. These actinometer-based protocols enable accurate light dose quantification for various scientific applications.

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Area of Science:

Biophysics
Microscopy
Photochemistry

Background:

Quantitative light intensity measurements are crucial in many scientific fields.
Current fluorescence microscopy techniques struggle to measure light dose with spatial resolution across diverse wavelengths and intensities.

Purpose of the Study:

To develop novel, rapid, and straightforward protocols for measuring light intensity and spatial distribution in fluorescence microscopy.
To address the limitations of existing technologies in quantifying light dose at the sample.

Main Methods:

Developed two protocols utilizing organic dyes and fluorescent proteins as actinometers.
Protocol 1: Employs molecular systems with monoexponential fluorescence decay/rise under constant light.
Protocol 2: Uses a photochemically inert, broad-absorbing fluorophore for inter-wavelength intensity back-calculation.

Main Results:

Successfully applied the protocols to quantitatively characterize light spatial distribution in various fluorescence imaging systems.
Demonstrated the calibration of illumination for commercial instruments and light sources.
Validated the accuracy and broad applicability of the developed actinometer-based methods.

Conclusions:

The developed actinometer protocols provide a robust solution for quantitative light dose measurement in fluorescence microscopy.
These methods enable precise characterization and calibration of illumination, improving experimental reproducibility across disciplines.
Offers a versatile tool for researchers needing accurate light intensity and spatial distribution data.