Performance comparison between the three unmixing algorithms (NNLS, NMF-SB and our NMF-ML) for the separation of FISH signals.

The median crosstalk coefficients (XT) are shown for each algorithm in synthetic and real images, together with the average calculation time for synthetic and real images. The best performer is highlighted in bold. As shown, the calculation time for the NMF-ML algorithm is drastically lower, especially in real images. This difference is mainly thanks to the ability to work on a 2D projection of the 3D stack.