Fig. 1: Fundamentals of single-molecule localization microscopy and illumination strategies.
a The principles of single-molecule super-resolution (SR) imaging. The localization of different emitters over time are used to form a SR reconstruction of the structure. b The principles of super-localization of emitters. The diffraction-limited image of the emitter is acquired, fitted by a model function, and subsequently super-localized. c The principles of single-molecule tracking. The localizations of the same emitter across multiple frames are linked to form a trajectory of the underlying dynamics. d The localization precision σ is shown as a function of signal photons per localization, Nsig, and background photons per pixel, Nbg, based on Eq. (1) using a 110 nm pixel size and 250 nm diffraction-limited spot size. e Comparison of different illumination strategies: wide-field epi-illumination (Epi), confocal illumination, total internal reflection fluorescence (TIRF) illumination, and light sheet illumination. Figures (a) and (c) are reprinted with permission from von Diezmann, et al. Chem. Rev. 117, 7244–7275 (2017). Copyright (2017) American Chemical Society.
