Spectral demixing (SD) offers multicolor single molecule localization microscopy (SMLM) with low crosstalk and without the need to correct for registration errors. Here, we present SDmixer, a versatile, open-source software tool that enables any laboratory to perform rapid SD-based multicolor SMLM. A graphic user interface allows non-experts to process 2D or 3D data sets from any SML software and to reconstruct the super-resolved multicolor images with flexible output options.