Unbiased quantitative ratiometric measurements in multiphoton microscopy with a generalized model

Emmanuelle Chaigneau; Marine Tournissac; Sonia Pfister; Mayeul Collot; Serge Charpak

doi:10.1117/12.2692652

12 March 2024 Unbiased quantitative ratiometric measurements in multiphoton microscopy with a generalized model

Emmanuelle Chaigneau, Marine Tournissac, Sonia Pfister, Mayeul Collot, Serge Charpak

Author Affiliations +

Proceedings Volume 12847, Multiphoton Microscopy in the Biomedical Sciences XXIV; 1284707 (2024) https://doi.org/10.1117/12.2692652
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

Fluorescent indicators allow the monitoring of physiological parameters in biological tissues by measuring changes in the brightness of the indicator upon binding of its ligand. Quantitative measurement of these parameters in vivo with high spatio-temporal resolution using multiphoton microscopy often requires the use of fluorescence ratiometric measurements. However, ratiometric measurements can be biased at depth in biological tissues due to absorption and scattering of the photons involved in the process.

Here we have developed a mathematical model that takes into account the chemical properties of the sensors, the spectral optical properties of the biological tissue and the optical system to provide quantitative measurements of the concentration of the sensor ligand. We have also developed a software implementing this model. Both can be used to obtain unbiased measurements of different physiological biomarkers from multiphoton images.

Conference Presentation

(2024) Published by SPIE. Downloading of the abstract is permitted for personal use only.

Citation Download Citation

Emmanuelle Chaigneau, Marine Tournissac, Sonia Pfister, Mayeul Collot, and Serge Charpak "Unbiased quantitative ratiometric measurements in multiphoton microscopy with a generalized model", Proc. SPIE 12847, Multiphoton Microscopy in the Biomedical Sciences XXIV, 1284707 (12 March 2024); https://doi.org/10.1117/12.2692652

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