Entering edit mode
2.1 years ago
mialopez97
•
0
I have a TSV file containing data that was previously processed by FlowJo, which consists of a column ID and several columns with parameters.
I have searched over the internet but I did not find the meaning of some of those parameters, like I know what the base parameters are, but there are some columns like Comp-FITC-A and Comp-PE-A that I do not know how are calculated.
If anyone has any resource about those compensations it will be much appreciated.
What do you mean base parameters?
These are compensated fluorescent channels. FITC and PE may be the channel nicknames for a certain excitation/emission combination, or they may be the actual fluorophores used in the experiment. "A" means the plots used the area data. The meaningful information though will be specific to the experiment in determing what marker occupies each channel. E.g. FITC may be the channel for a LysM-GFP reporter, or something else....
The compensation is a calculation done to extract the bleed over between channels. Simply, the software takes single-color controls and calculates the bleed over into other channels. (e.g. take FITC-only cells and see if there is signal in PE, or vice versa). Once these are calculated the software takes co-stained/multi-color samples and subtracts the same % of bleed over determined by the compensation controls.
You can probably use these columns the same as if they were simply labeled "FITC-A" and "PE-A".