Speaker
Description
In this workshop we propose to learn the principles of phasor analysis of Fluoresce Lifetime Microscopy (FLIM) data as well as to perform image analysis of FLIM data with this approach using (F)luorescence (L)ifetime (U)l(t)imate (E)xplorer (FLUTE) a new open source and user-friendly GUI that we recently developed [1,2]. Using FLUTE, participants will learn the basis of FLIM calibration, phasor plotting, FLIM data interactive visualization and exploration, data interpretations and advanced phasor analysis and quantification. We also propose to explore and understand the impact of the most important parameters and factors of FLIM acquisition, such as the number of photons, as well as sources of bias such as the presence of background. The phasor analysis will be performed on a variety a FLIM data acquired in the time domain from different fluorophores, autofluorescence from zebrafish embryo and cells in culture. Example of functional metabolic images will be explored. It will also be possible to analyze FLIM data that users will bring from their labs and/or acquired with different FLIM commercial systems.
[1] D Gottlieb, B Asadipour, P. Kostina, TPL Ung, C Stringari. FLUTE: a Python GUI for interactive phasor analysis of FLIM data. Gottlieb D, Asadipour B, Kostina P, Ung TPL, Stringari C. FLUTE: A Python GUI for interactive phasor analysis of FLIM data. Biological Imaging. 2023;3:e21.
[2] https://github.com/LaboratoryOpticsBiosciences/FLUTE
| Target audience | Beginner users, Intermediate users, Advanced users, Intermediate developers |
|---|---|
| Keywords | Data visualization, fluorescence lifetime microscopy, FLIM, phasor analysis, Python |
