Super-resolution tracking of weak fluorescent markers in 3D: application in dissecting the mechanics of chromosome segregation using fluorescence imaging and molecular genetics in yeast

We present a framework for the tracking of fluorescent tags in 4D microscopy (3D + time). It includes algorithms to separate tag signals under strong interference, allowing us to break the limits of optical resolution in light microscopy by a factor 3. We apply the package to read out chromosome dynamics in a functional, cell-based screen for proteins regulating cell division in yeast.