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Related Experiment Video

Updated: Feb 23, 2026

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy

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32-channel time-correlated-single-photon-counting system for high-throughput lifetime imaging.

P Peronio¹, I Labanca¹, G Acconcia¹

¹Dipartimento di Elettronica, Informazione e Bioingegneria, Politecnico di Milano, Piazza Leonardo da Vinci 32, 20133 Milano, Italy.

The Review of Scientific Instruments

|September 3, 2017

Summary

This study presents an advanced multichannel Time-Correlated Single Photon Counting (TCSPC) system to overcome measurement time limitations in optical signal detection. The new system enhances performance for high-throughput fluorescence lifetime imaging microscopy.

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Last Updated: Feb 23, 2026

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Area of Science:

Photonics and Optical Instrumentation
Biophysics
Microscopy

Background:

Time-Correlated Single Photon Counting (TCSPC) is crucial for fast, weak optical signal measurement but suffers from long acquisition times.
Existing multichannel TCSPC systems often face a performance trade-off with increasing channel parallelism.
A previous 32-channel TCSPC system demonstrated overcoming this trade-off.

Purpose of the Study:

To develop an advanced TCSPC system for high-throughput fluorescence lifetime imaging microscopy (FLIM).
To overcome the performance limitations of existing multichannel TCSPC systems.
To enable faster and more efficient FLIM acquisition.

Main Methods:

Design and implementation of a novel hardware architecture for a multichannel TCSPC system.
Development of centralized logic and fast data management firmware.
Integration with a microscope for practical FLIM applications.

Main Results:

The new system architecture successfully addresses the trade-off between parallelism and performance in TCSPC.
Demonstrated feasibility for high-throughput FLIM applications.
Preliminary results validate the system's performance and applicability.

Conclusions:

The evolved multichannel TCSPC system offers a viable solution for high-throughput FLIM.
The system's design overcomes previous performance limitations in parallel TCSPC.
This technology advances optical signal measurement capabilities for advanced microscopy.