Pixelation with concentration-encoded effective photons for molecular optical sectioning microscopy

Geng Wang; Rishyashring Iyer; Janet Sorrells; Edita Aksamitiene; Eric Chaney; Carlos Renteria; Jaena Park; Jindou Shi; Yi Sun; Stephen Boppart; Haohua Tu

doi:10.1117/12.3001809

13 March 2024 Pixelation with concentration-encoded effective photons for molecular optical sectioning microscopy

Geng Wang, Rishyashring Iyer, Janet Sorrells, Edita Aksamitiene, Eric Chaney, Carlos Renteria, Jaena Park, Jindou Shi, Yi Sun, Stephen Boppart, Haohua Tu

Author Affiliations +

Proceedings Volume PC12836, Optical Biopsy XXII: Toward Real-Time Spectroscopic Imaging and Diagnosis; PC128360H (2024) https://doi.org/10.1117/12.3001809
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

Quality control in molecular optical sectioning microscopy is indispensable for transforming acquired digital images from qualitative descriptions to quantitative data. Although numerous tools, metrics, and phantoms have been developed, accurate quantitative comparisons of data from different microscopy systems with diverse acquisition conditions remains a challenge. Here, we develop a simple tool based on an absolute measurement of bulk fluorophore solutions with related Poisson photon statistics, to overcome this obstacle. Demonstrated in a prototypical multiphoton microscope, our tool unifies the unit of pixelated measurement to enable objective comparison of imaging performance across different modalities, microscopes, components/settings, and molecular targets. The application of this tool in live specimens identifies an attractive methodology for quantitative imaging, which rapidly acquires low signal-to-noise frames with either gentle illumination or low-concentration fluorescence labeling.

Conference Presentation

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

Citation Download Citation

Geng Wang, Rishyashring Iyer, Janet Sorrells, Edita Aksamitiene, Eric Chaney, Carlos Renteria, Jaena Park, Jindou Shi, Yi Sun, Stephen Boppart, and Haohua Tu "Pixelation with concentration-encoded effective photons for molecular optical sectioning microscopy", Proc. SPIE PC12836, Optical Biopsy XXII: Toward Real-Time Spectroscopic Imaging and Diagnosis, PC128360H (13 March 2024); https://doi.org/10.1117/12.3001809

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KEYWORDS

Photons

Optical microscopy

Microscopy

Multiphoton fluorescence microscopy

Fluorescence

Fluorescence microscopy

Reproducibility

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