Fluorescence-lifetime imaging microscopy (FLIM) not only for biologists - practical aspects with hands-on experience - MB100P03
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Last update: Mgr. Aleš Benda, Ph.D. (23.09.2020)
The course focuses on explaining the principles of nano-second time-resolved fluorescence detection, on demonstration of different hardware realizations, on trying various applications of FLIM in biological imaging and on testing of several ways of FLIM data analysis. The aim is to uncover the richness of information hidden in multi-parametric fluorescence imaging and to inspire the participants to use the easily obtainable extra contrast in their imaging applications. An important part is to make the FLIM data analysis understandable. The course will cover the most widespread FLIM realization – acquisition in time domain based on raster scanning combined with pulsed lasers and time-correlated single photon counting (TCSPC). Participants will hands-on three different systems, covering pulsed diode lasers, white-light laser source as well as two-photon excitation, SPAD, HyD and GaAsP NDD detectors and three different TCSPC systems and software. A focus will be given on common pitfalls, instrumental artifacts and ways of avoiding them. The demonstrated applications will include FLIM-FRET, environmental sensing, metabolic NAD(P)H imaging and FLIM data analysis. |
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Last update: Mgr. Aleš Benda, Ph.D. (08.06.2022)
The bible of FLIM is the handbook from Wolfgang Becker: https://www.becker-hickl.com/literature/documents/flim/the-bh-tcspc-handbook/ Technical aspects of the main applications are described here: https://www.tcspc.com/doku.php/applications#fluorescence_lifetime_imaging_microscopy_flim A recent review on newest applications, including those demonstrated during the course, is J. of Biomedical Optics, 25(7), 071203 (2020). https://doi.org/10.1117/1.JBO.25.7.071203 |
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Last update: Mgr. Aleš Benda, Ph.D. (07.10.2019)
Active participation at the whole course plus short 3 min presentation of your own work or planned work.. |
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Last update: Mgr. Aleš Benda, Ph.D. (23.09.2020)
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Last update: Mgr. Aleš Benda, Ph.D. (12.09.2018)
The course is intended for magister and doctoral students. |