New light microscope for highest precision in single-molecule localisation and tracking

A limited photon budget, together with sample drift, are the main factors that prevent other nanoscopy techniques from reaching their theoretical potential. Like other single molecule localisation techniques, such as STORM/PALM, MINFLUX takes advantage of the on-off transition of fluorescent molecules to separate the emission of densely packed emitters over time. Unlike the aforementioned camera-based techniques, MINFLUX follows a hybrid coordinate-targeted/stochastic approach: localisation of each individual fluorophore is achieved by “injecting” a reference coordinate in the sample in the form of a local excitation minimum, a strategy derived from Stimulated Emission Depletion (STED) microscopy. The superior photon efficiency of the MINFLUX technology, being 20 times higher than camera-based techniques, in combination with its very effective active sample stabilisation system, are responsible for its unprecedented localisation precision.