The data and information in this repository is related to a workshop on Fluorescence Lifetime Imaging Microscopy for the simultaneous imaging of two or three spectrally identical fluorescent proteins. The workshop "Atélier 4" was done twice on November 12th, 2023 for the MiFoBio meeting: https://imabio-cnrs.fr/mifobio/programme-2023/ This repository includes: a pdf file with the description & planning of the workshop a ppt file with brief introductory presentation to introduce the topic raw data (as ptu file) that was acquired in preparation of the workshop, on the same samples that were used in the workshop, using similar equipment. processed data as tif file The example FLIM data were acquired with a Leica Stellaris8 microscope at the The Van Leeuwenhoek Centre for Advanced Microscopy in Amsterdam: http://www.lcam-fnwi.nl/ The plasmids used in the workshop are available on addgene (https://www.addgene.org/60491/) or will be made available (please contact me if you cannot find the plasmid info) in the near future. The .ptu file contain the raw data, the _ij.tif file show the lifetime data based on mean arrival times and the _fractions.tif show the unmixed images.