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Updated: Nov 9, 2025

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

Simultaneous multi-spectral, single-photon fluorescence imaging using a plasmonic colour filter array.

Peter W R Connolly¹, Jessica Valli¹, Yash D Shah^2,3

¹School of Engineering and Physical Sciences, Heriot-Watt University, Edinburgh, UK.

Journal of Biophotonics

|April 8, 2021

Summary

This summary is machine-generated.

Researchers developed a new method for simultaneous multi-spectral fluorescence imaging using a single-photon avalanche diode (SPAD) array and a plasmonic metasurface color filter array (CFA). This innovation enables high-fidelity, full-color imaging with minimal light, paving the way for real-time applications.

Keywords:

CFA FLIM SPAD colour filter array fluorescence image processing imaging metasurface microscopy plasmonic single-photon avalanche diode

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Last Updated: Nov 9, 2025

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Published on: September 4, 2013

Area of Science:

Optics and Photonics
Biomedical Imaging
Materials Science

Background:

Multi-spectral fluorescence imaging is crucial for biological research, but often requires complex and bulky equipment.
Existing methods can be slow, require high light levels, or necessitate multiple detectors.
Single-photon avalanche diode (SPAD) arrays offer high sensitivity but integrating spectral information has been challenging.

Purpose of the Study:

To demonstrate the first simultaneous multi-spectral fluorescence imaging system using a SPAD array.
To develop a compact and efficient spectral unmixing method for fluorescence imaging.
To enable high-fidelity, full-color imaging with low photon counts.

Main Methods:

Utilized a 64x64 pixel silicon SPAD array for data acquisition.
Employed a plasmonic metasurface mosaic color filter array (CFA) for spectral separation.
Developed a specialized spectral unmixing algorithm for image reconstruction.

Main Results:

Achieved simultaneous multi-spectral fluorescence imaging with high fidelity and full color.
Reconstructed high-quality images from as few as approximately 3 photons per pixel.
Demonstrated the system's capability with four biological samples.

Conclusions:

The developed system offers a novel, compact solution for multi-spectral fluorescence imaging.
This approach facilitates real-time, single-photon sensitive imaging without bulky optical components.
The technology holds promise for advancing biological research and diagnostics.