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Updated: Jun 3, 2025

Single Molecule Fluorescence Microscopy on Planar Supported Bilayers

Single Molecule Fluorescence Microscopy on Planar Supported Bilayers

Published on: October 31, 2015

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

Geng Wang^1,2, Rishyashring R Iyer^1,2, Janet E Sorrells^1,3

¹Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.

Laser & Photonics Reviews

|January 9, 2025

Summary

A new tool quantifies molecular optical sectioning microscopy images using absolute fluorophore measurements. This enables objective comparisons across diverse microscopy systems and settings, advancing quantitative biological imaging.

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Area of Science:

Biophotonics
Microscopy
Quantitative Imaging

Background:

Irreproducibility in molecular optical sectioning microscopy limits quantitative data acquisition.
Existing tools struggle with accurate comparisons across different systems and conditions.

Purpose of the Study:

To develop a simple, objective tool for quantifying microscopy performance.
To enable accurate comparisons of imaging data across diverse modalities and settings.

Main Methods:

Utilized absolute measurements of bulk fluorophore solutions.
Incorporated Poisson photon statistics for analysis.
Demonstrated on a multiphoton microscope.

Main Results:

Unified pixelated measurements into a common unit.
Enabled objective comparison of imaging performance.
Validated for live specimen imaging.

Conclusions:

The developed tool overcomes challenges in quantitative microscopy comparisons.
Facilitates rapid acquisition of low signal-to-noise images for quantitative analysis.
Presents an attractive methodology for quantitative imaging in biological specimens.