Compared to conventional compensation, spectral unmixing distinguishes the shapes of emission spectra along a large range of wave lengths. This approach can discriminate fluorochromes of similar emission spectrum, as well as provide better separation. Although the measurement of a continuous wave length is impossible in conventional cytometry, a high parameter cytometer can well cover most of the light spectrum by enabling all detectors. This provides a “pseudo-spectral approach” that can provide better signal separation than conventional compensation in theory. This presentation will discuss how high parameter data may benefit from utilizing pseudo spectral unmixing instead of normal compensation. Procedures for spectral unmixing on FlowJo V10.6 will also be covered.