Authors
Emmanuelle Chaigneau, Marine Tournissac, Sophie Walter, Mayeul Collot, Serge Charpak
Published in
Neurophotonics. Volume 13. Issue 3. Pages 035007. Epub Sep 03, 2026.
Abstract
Multiphoton microscopy with fluorescent indicators enables the monitoring of biomarker dynamics through changes in fluorescence intensity. However, quantitative interpretation remains challenging because fluorescence signals are highly sensitive to experimental conditions. Dual-color ratiometric approaches improve quantification, yet wavelength-dependent absorption and scattering of emitted photons can still introduce significant bias.
This study aimed to develop a correction method for absorption and scattering effects to achieve unbiased dual-color ratiometric measurements in multiphoton microscopy.
We developed an analytical model that incorporates the spectral optical properties of biological tissues and fluorophores. The model was applied to the rodent cerebral cortex parenchyma and to the lumen of individual blood vessels. Its predictions were evaluated using simulations and in vivo two-photon microscopy experiments.
In the cerebral cortex parenchyma, absorption and scattering bias dual-color ratiometric measurements by up to at depths of approximately . Our model accurately describes and corrects these effects. Within the vessel lumen, however, the situation is more complex: the model effectively corrects the biases in veins, but not in arterioles.
Overall, the proposed model enhances the accuracy of biomarker quantification in multiphoton microscopy and provides practical tools for correcting optical biases in biological tissues.
PMID:
42694932
Bibliographic data and abstract were imported from PubMed on 04 Sep 2026.
Read full publication at:
Please sign in
to see all details.
Advertisement
Stats
- Recommendations n/a n/a positive of 0 vote(s)
- Views 6
- Comments 0