The Bright-Eyes TTM: A time-tagging module to democratize single-photon microscopy
Overview
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. 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.
Speaker
Eleonora Perego studied Physics at the University of Milano-Bicocca (Italy). She got a M. Sc. in Physics in 2015, after a period at the AMOLF institute in Amsterdam (the Netherlands) with a thesis titled “The role of mechanical forces in the robustness of C. elegans embryonic development”. She joined for her PhD the “Cellular Biophysics” group of Prof. Dr. Sarah Koester (Göttingen, Germany), where she studied with fluorescence spectroscopy methods combined with microfluidic approaches protein-protein interactions. In 2020, Eleonora obtained her PhD in Biophysics at the Göttingen Graduate School for Neurosciences Biophysics and Molecular Biosciences (GGNB, Göttingen, Germany) under the program of the International Max Planck Research School “Physics of Biological and Complex Systems”, with a thesis titled “Studying molecular interaction under flow with fluorescence fluctuation spectroscopy”. Since 2021, Eleonora is working at the Italian Institute of Technology in Genoa, Italy, in the Molecular Microscopy and Spectroscopy research line of Dr. G. Vicidomini as a post-doctoral researcher. Her research interests include the application of fluorescence spectroscopy methods, such as fluorescence fluctuation spectroscopy and fluorescence lifetime imaging, to biological questions, ranging from neuroscience to RNA biology.
Eli Slenders has a bachelor degree in Physics from Hasselt University (Belgium) and graduated in 2013 as a master of Science in Physics from KU Leuven (Belgium). In 2018, Eli obtained his PhD in Physics from Hasselt University under supervision of Prof. Dr. M. Ameloot with a thesis entitled “Resolution in coherent and incoherent optical imaging with two-photon excitation microscopy”. Since 2019, Eli is employed at the Italian Institute of Technology in Genoa, Italy, in the Molecular Microscopy and Spectroscopy research line of Dr. G. Vicidomini, first as a post-doctoral researcher, currently as a Marie Skłodowska-Curie fellow. His research interests include the theoretical design and experimental validation of optical microscopy tools such as multiphoton image scanning microscopy and fluorescence correlation spectroscopy. His current work focuses on single-molecule imaging.
Related Publications
Rossetta A, Slenders E, Donato M, Zappone S, Fersini F, Bruno M, Diotalevi F, Lanzanò L, Koho S, Tortarolo G, Barberis A, Crepaldi M, Perego E, Vicidomini G. The BrightEyes-TTM as an open-source time-tagging module for democratising single-photon microscopy. Nat Commun. 2022 Dec 1;13(1):7406. doi: 10.1038/s41467-022-35064-0. PMID: 36456575; PMCID: PMC9715684.
Ranjit S, Malacrida L, Jameson DM, Gratton E. Fit-free analysis of fluorescence lifetime imaging data using the phasor approach. Nat Protoc. 2018 Sep;13(9):1979-2004. doi: 10.1038/s41596-018-0026-5. PMID: 30190551.
Slenders E, Perego E, Buttafava M, Tortarolo G, Conca E, Zappone S, Pierzynska-Mach A, Villa F, Petrini EM, Barberis A, Tosi A, Vicidomini G. Cooled SPAD array detector for low light-dose fluorescence laser scanning microscopy. Biophys Rep (N Y). 2021 Dec 8;1(2):None. doi: 10.1016/j.bpr.2021.100025. PMID: 34939046; PMCID: PMC8651514.
Slenders, E., Castello, M., Buttafava, M. et al. Confocal-based fluorescence fluctuation spectroscopy with a SPAD array detector. Light Sci Appl 10, 31 (2021). https://doi.org/10.1038/s41377-021-00475-z
Repositories
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