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Related Experiment Video

Updated: May 19, 2026

Local Field Fluorescence Microscopy: Imaging Cellular Signals in Intact Hearts

Local Field Fluorescence Microscopy: Imaging Cellular Signals in Intact Hearts

Published on: March 8, 2017

Counting fluorescent emitters with a single photon avalanche diode array.

Clayton Seitz, Carmella Evans-Molina, Jing Liu

Biorxiv : the Preprint Server for Biology

|May 18, 2026

Summary

This summary is machine-generated.

Photon counting histogram (PCH) analysis is now possible in widefield microscopy using single photon avalanche diode (SPAD) arrays. This breakthrough enables precise quantification of active fluorophore numbers, advancing fluorescence imaging techniques.

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Last Updated: May 19, 2026

Local Field Fluorescence Microscopy: Imaging Cellular Signals in Intact Hearts

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Published on: March 8, 2017

Fluorescence detection methods for microfluidic droplet platforms

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Published on: December 10, 2011

Advanced Compositional Analysis of Nanoparticle-polymer Composites Using Direct Fluorescence Imaging

Advanced Compositional Analysis of Nanoparticle-polymer Composites Using Direct Fluorescence Imaging

Published on: July 19, 2016

Area of Science:

Fluorescence Microscopy
Quantum Optics
Biophysics

Background:

Photon counting histogram (PCH) traditionally quantifies fluorophores in single focal volumes.
Widefield imaging lacked methods to capture single fluorophore emission statistics due to sensor limitations.

Purpose of the Study:

To adapt PCH analysis for widefield fluorescence microscopy.
To quantify active fluorophore numbers using advanced SPAD array technology.

Main Methods:

Utilizing single photon avalanche diode (SPAD) arrays synchronized with picosecond laser pulses.
Measuring PCH in a widefield setting via binary exposures.
Applying Bayesian inference to model the relationship between fluorophore number and PCH.

Main Results:

Successfully demonstrated PCH measurement in a widefield setup.
Quantified numbers of quantum dots and DNA-bound dye molecules.
Validated the Bayesian inference model for fluorophore counting.

Conclusions:

SPAD array technology enables widefield PCH analysis for the first time.
This method offers accurate quantification of active fluorophore numbers.
Applications include enhanced localization microscopy and fitting unresolvable emitters.