The BrightEyes-TTM as an open-source time-tagging module for democratising single-photon microscopy
Alessandro Rossetta, Eli Slenders, Mattia Donato, Sabrina Zappone, Francesco Fersini, Martina Bruno, Francesco Diotalevi, Luca Lanzanò, Sami Koho, Giorgio Tortarolo, Andrea Barberis, Marco Crepaldi, Eleonora Perego, Giuseppe Vicidomini
Abstract
Fluorescence laser-scanning microscopy (LSM) is experiencing a revolution thanks to new single-photon (SP) array detectors, which give access to an entirely new set of single-photon information. Together with the blooming of new SP LSM techniques and the development of tailored SP array detectors, there is a growing need for (i) DAQ systems capable of handling the high-throughput and high-resolution photon information generated by these detectors, and (ii) incorporating these DAQ protocols in existing fluorescence LSMs. We developed an open-source, low-cost, multi-channel time-tagging module (TTM) based on a field-programmable gate array that can tag in parallel multiple single-photon events, with 30 ps precision, and multiple synchronisation events, with 4 ns precision. We use the TTM to demonstrate live-cell super-resolved fluorescence lifetime image scanning microscopy and fluorescence lifetime fluctuation spectroscopy. We expect that our BrightEyes-TTM will support the microscopy community in spreading SP-LSM in many life science laboratories.