The BrightEyes-TTM as an open-source time-tagging module for democratising single-photon microscopy

Area of Science:

• Optics and Photonics
• Biophysics
• Microscopy Technology

Background:

• Single-photon (SP) array detectors are revolutionizing fluorescence laser-scanning microscopy (LSM).
• Existing data acquisition (DAQ) systems struggle with the high-throughput, high-resolution photon data from new SP detectors.
• There is a need for integrated DAQ solutions for advanced SP LSM techniques.

Purpose of the Study:

• To develop an open-source, low-cost data acquisition system for SP array detectors in LSM.
• To enable high-precision time-tagging of single-photon and synchronization events.
• To facilitate the integration of advanced SP LSM techniques into existing microscopy setups.

Main Methods:

• Development of a multi-channel time-tagging module (TTM) using a field-programmable gate array (FPGA).
• Achieved 30 ps precision for single-photon event tagging and 4 ns precision for synchronization events.
• Integration of the TTM with fluorescence LSM systems.

Main Results:

• Demonstrated the TTM's capability for high-throughput, high-resolution photon data acquisition.
• Successfully applied the TTM to live-cell super-resolved fluorescence lifetime image scanning microscopy.
• Utilized the TTM for fluorescence lifetime fluctuation spectroscopy.

Conclusions:

• The developed BrightEyes-TTM provides a crucial DAQ solution for the growing field of SP LSM.
• This open-source module supports the widespread adoption of SP LSM in life science research.
• The TTM enhances capabilities for advanced live-cell imaging and spectroscopic analysis.