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Basic working principle of STED nanoscopy. (a) Focal spot of conventional confocal microscopy. Fluorophores in the PSF of the excitation laser undergo fluorescent transitions. (b) Focal spot of the STED nanoscopy. Only fluorophores at the very center of the STED beam undergo fluorescent transitions. (c) Jablonski diagram of two molecular transitions: fluorescent transition (spontaneous emission) and nonfluorescent transition (stimulated emission).
