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Updated: Jan 19, 2026

Binet's Contribution to Measures of Intelligence

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Molecular contribution function in RESOLFT nanoscopy.

Lars Frahm, Jan Keller-Findeisen, Philipp Alt

|September 13, 2019

Summary

This summary is machine-generated.

This study introduces a new method using RESOLFT nanoscopy to count molecules by analyzing the molecular contribution function (MCF). This approach enables precise molecular counting within subdiffraction-sized volumes for advanced microscopy applications.

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Molecular Evolution of the Tre Recombinase

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Published on: May 29, 2008

Area of Science:

Microscopy
Biophysics
Molecular Imaging

Background:

High-resolution microscopy aims to map molecules within samples.
Traditional imaging relies on the point spread function (PSF) to characterize spatial resolution.
Existing methods face challenges in direct molecular counting.

Purpose of the Study:

To develop a quantitative method for counting fluorophores using RESOLFT nanoscopy.
To extend the concept of PSF to the molecular contribution function (MCF).
To enable precise molecular counting within subdiffraction-sized regions.

Main Methods:

Utilized reversible saturable optical fluorescence transitions (RESOLFT) for targeted fluorophore activation.
Developed a two-state Markov model to approximate the MCF, mimicking rsEGFP2 switching kinetics.
Analyzed signal variance and mean during readout to quantify fluorophore contribution.

Main Results:

Successfully approximated the molecular contribution function (MCF) for RESOLFT nanoscopy.
Demonstrated that signal variance scales quadratically with fluorophore brightness, while the mean scales linearly.
Validated the method on tubulin structures in Drosophila melanogaster larvae, enabling accurate fluorophore counting.

Conclusions:

Modeling and estimation of the MCF provide a quantitative approach for RESOLFT nanoscopy.
This method offers a straightforward concept for counting fluorophores at the molecular level.
The developed technique is a promising advancement for quantitative molecular imaging and microscopy.