Ostersehlt, Lynn M.Lynn M.OstersehltJans, Daniel C.Daniel C.JansWittek, AnnaAnnaWittekKeller-Findeisen, JanJanKeller-FindeisenInamdar, KaushikKaushikInamdarSahl, Steffen J.Steffen J.SahlHell, Stefan W.Stefan W.HellJakobs, StefanStefanJakobs2023-06-072023-06-072022https://publica.fraunhofer.de/handle/publica/44261910.1038/s41592-022-01577-12-s2.0-8513706672436050490MINimal fluorescence photon FLUXes (MINFLUX) nanoscopy, providing photon-efficient fluorophore localizations, has brought about three-dimensional resolution at nanometer scales. However, by using an intrinsic on-off switching process for single fluorophore separation, initial MINFLUX implementations have been limited to two color channels. Here we show that MINFLUX can be effectively combined with sequentially multiplexed DNA-based labeling (DNA-PAINT), expanding MINFLUX nanoscopy to multiple molecular targets. Our method is exemplified with three-color recordings of mitochondria in human cells.enjournal article