flowjo compensation matrix negative values

A size gate that removes outliers is still essential for isolating representative events, and the size gate drawn by FlowJo may need adjustment to contain the representative populations.

Unlike traditional compensation, the AutoSpill method does not require gated positive and negative populations, as the algorithm will determine its own positive populations. FlowJo may also try to load an unstained control for use as a universal negative. Compensation controls must first be loaded into the FlowJo compensation group, then the compensation button at the top of the workspace can be selected.įlowJo will automatically try to match the compensation controls to the appropriate detectors, apply a size gate to each of the controls, then try to gate the positive and negative populations in each control. Performing AutoSpill compensation has similar setup to regular compensation. The option to return to traditional compensation is located next to the AutoSpill tick box and there is also a button for removing the universal negative, which is not required for the AutoSpill algorithm but can be used to remove autofluorescence. When the option is ticked FlowJo will use the AutoSpill algorithm to calculate the compensation matrix. The AutoSpill option can be found in the compensation wizard. Additionally the AutoSpill function can be used to remove the signal due to autofluorescence. The new algorithm, combines automatic gating with linear regression through an iterative process that helps reduce error and improve the compensation matrix. This form of gate-less compensation removes some of the requirements of traditional compensation making compensation faster and simpler. AutoSpill is a new algorithm for calculating spillover and producing a compensation matrix developed and incorporated into FlowJo v10.7 in collaboration with researchers from the Vlaams Instituut voor Biotechnologie in Belgium and the the Babraham Institute in Cambridge, UK.

Flowjo compensation matrix negative values