Authors

Molecular Microscopy & Spectroscopy, Istituto Italiano di Tecnologia

and

Giuseppe Vicidomini [1],[0],[*]

Contributions

G.V. conceived the idea and designed the research;
G.V. supervised the research;
A.R., M.D., with the support from F.D. and M.C., wrote the firmware for the time-tagging module and the data-receiver;
M.D., with support from A.R., wrote the operational software;
A.R., E.S., M.D., S.K., and G.T. wrote the imaging analysis software;
E.S., M.D., and L.L. wrote the FFS analysis software;
A.R. and M.D. characterized and validated the TTM module with the test-bench architecture;
E.S. and F.F. built the single-photon microscopes;
S.Z. prepared the cell samples. E.S. and E.P., with support from A.R., performed the single-photon microscopy experiments;
E.P. designed, implemented, and analyzed all the FLFS experiments;
A.R., E.S., M.D., E.P., S.K., G.T., and G.V., analyzed the data;
A.R., M.D., and G.V. built the GitHub repository;
G.V., E.S. and E.P., with support from A.R. and M.D., wrote the manuscript with edits from all authors.

Acknowledgements

This research was supported by Fondazione San Paolo, “Observation of bio-molecular processes in live-cell with nanocamera”, No. EPFD0098 (E.S. and G.V.), by the European Research Council, Bright Eyes, No. 818699 (G.T. and G.V.), and by the European Union’s Horizon 2020 research and innovation programme under the Marie Sk1odowska-Curie grant agreement no. 890923 (SMSPAD) (E.S.). We thank Prof. Alberto Diaspro and Dr. Paolo Bianchini (Nanoscopy & NIC@IIT, Istituto Italiano di Tecnologia) for useful discussions; Dr. Michele Oneto (Nikon Imaging Center) and Marco Scotto (Molecular Microscopy and Spectroscopy, Istituto Italiano di Tecnologia) for support on the experiments; Dr. Andrea Barberis and Dr. Enrica Petrini (Synaptic Plasticity of Inhibitory Networks, Istituto Italiano di Tecnologia) for providing the primary hippocampal cells; Alessandro Barcellona (Electronic Design Laboratory, Istituto Italiano di Tecnologia) for design and implementation of the custom-made buffer; Prof. Alberto Tosi, Prof. Federica Villa, Dr. Mauro Buttafava (Politecnico di Milano), Dr. Marco Castello, and Dr. Simonluca Piazza (Istituto Italiano di Tecnologia and Genoa Instruments) for useful initial discussions in the time-to-digital design and for the realization of the single-photon-avalanche-diode detector array.

Copyright

License

The Time-Tagging Module (TTM) is an aggregation of different parts with different licenses. See details in LICENSE.md file. Unless otherwise stated, they are licensed under a CC-BY-NC 4.0, Creative Commons Attribution-NonCommercial 4.0 International License.

In addition to the terms of the license, we ask to acknowledge the use of the time-tagging module in scientific articles by citing:

Rossetta, A., Slenders, E., Donato, M. et al.
The BrightEyes-TTM as an open-source time-tagging module for democratising single-photon microscopy. 
Nat Commun 13, 7406 (2022). 
https://doi.org/10.1038/s41467-022-35064-0

Contact us

Do you need help to build up your time-tagging module? Do you have comments or questions? Do not hesitate to contact us at mailto:giuseppe.vicidomini@iit.it.

References

[1] A. Rossetta, E. Slenders, M. Donato, E. Perego, F. Diotalevi, L. Lanzano’, S. V. Koho, G. Tortarolo, M. Crepaldi, G. Vicidomini bioRxiv 2021.10.11.463950; doi: https://doi.org/10.1101/2021.10.11.463950

[2] M. Castello, G. Tortarolo, M. Buttafava, T. Deguchi, F. Villa, S. Koho, L. Pesce, M. Oneto, S. Pelicci, L. Lanzanó, P. Bianchini, C. J. R. Sheppard, A. Diaspro, A. Tosi, and G. Vicidomini, A robust and versatile platform for image scanning microscopy enabling super-resolution FLIM, Nat. Methods , 16: 175–178 (2019), https://doi.org/10.1038/s41592-018-0291-9

[3] E. Slenders, M. Castello, M. Buttafava, F. Villa, A. Tosi, L. Lanzano, S. V. Koho, and G. Vicidomini, 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

[4] M. Buttafava, F. Villa, M. Castello, G. Tortarolo, E. Conca, M. Sanzaro, S. Piazza, P. Bianchini, A. Diaspro, F. Zappa, G. Vicidomini, and A. Tosi, SPAD-based asynchronous-readout array detectors for image-scanning microscopy, Optica , 7: 755-765 (2020), https://doi.org/10.1364/OPTICA.391726

[5] S. V. Koho, E. Slenders, G. Tortarolo, M. Castello, M. Buttafava, F. Villa, E. Tcarenkova, M. Ameloot, P. Bianchini, C. J. R. Sheppard, A. Diaspro, A. Tosi, and G. Vicidomini, Two-photon image-scanning microscopy with SPAD array and blind image reconstruction, Biomed. Opt. Express , 11(6): 2905-2924 (2020), https://doi.org/10.1364/BOE.374398