Evaluating MINFLUX experimental performance in silico

MINFLUX is a super-resolution microscopy technique with remarkable resolution for imaging and tracking. Its optical resolution is well understood under idealized conditions, but because of system complexity and experimental imperfections, actual performance can be much worse. Here we present SimuFLUX, a comprehensive and realistic simulator for MINFLUX. We use it to investigate performance limits caused by fluorophore dynamics, background, estimators, and misalignment, and to simulate experiments to optimize parameters and assess feasibility.