The matrix is then inverted and gives the actual compensation values. This is done by measuring the spectral overlap of the different fluorochromes and using the measured values to create a matrix. This correction is called compensation.Ĭompensation is necessary in order to be able to differentiate between populations of cells. The ability to correct this stems from the fact, that the overlap is a linear function, so the measured signal can be averaged and thus corrected. The physical overlap between the different emission spectra of fluorochromes can activate different receptors than the ones intended for the given wavelength. When one cell is marked by two or more fluorochromes, the added brightness of one fluorochrome to the other creates significant background noise and affects the strength of the signal. during a two colour experiment, where mouse splenocytes were stained with fluorescein and rhodamine. The first data compensation was done in 1977 by Michael Loken et al. The compensation can be done through different flow cytometry software such as Flowjo, Flowlogic, Kaluza etc. This creates a signal overlap (spillover) which cannot be removed by the optical system and has to be corrected electronically. The photons emitted by fluorochromes have different energies and wavelengths and as flow cytometers use photomultiplier tubes (PMT) in order to convert the photons into electrons, the detector can register the signal from more than one fluorochrome. This service is currently free of charge, too.In cytometry, compensation is a mathematical correction of a signal overlap between the channels of the emission spectra of different fluorochromes. If you are sorting in the CFFC you will receive step-by-step tuition and guidance leading you to being able to work with the FACSAria sorter independently and operator-free. This workshop is co-organized by TransMed and the Core Facility Flow Cytometry (CFFC)Ībout the Core Facility Flow Cytometry (CFFC)Įveryone is welcome to use FlowJo TM on the CFFC workstations, currently free of charge. Please bring your own data with you in order to analyze them.įor the workshop 1 CP for scientific skills training can be credited (0.5 CP for participating only in the theoretical part). Please read the FlowJo TM description, which will be sent to you after your registration. Theoretical and practical input by the trainer, showing examples, analyzing own data using FlowJo TM with the help of the trainer, discussing own issues, feedback from the trainer.ĭoctoral students, postdocs and clinician scientists who already know the basics of FlowJo TM.
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