STEDSTED = STimulated-Emission-DepletionBasic photophysicsLeutenegger et al. (2010) Optics Express 18, 26417-26429.S1*S1Principles of STEDYamanaa et al. (2014) !icr"sc"p# 63, 177-192. $lar et al. (2000) %&'( 97, )206-)210.Principles of STEDYamanaa et al. (2014) !icr"sc"p# 63, 177-192. $lar et al. (2000) %&'( 97, )206-)210.*ell (2007) (cience 316, 11+,-11+).Factors that Affect Spatial Resolution IS: intensity that maes stimulated emission rate e!ual spontaneous decay rate Ima"#IS cannot $e made ar$itrarily lar%e Photo$leachin% Dama%e to sample Finite spontaneous decay rate &a"imal spatial resolution of $iolo%ical samples:'()nm *until +),(-./Laa-am#ali. (2014) ./em%/#s./em 15, 6,0-6,6.0enerali1ation: RES23FTRE4ersi$le Satura$le 2ptical F3uorescence Transitions Super-resolution microscopy that in4ol4es saturation#depolari1ation of dar#fluorescent state 0enerali1ation of STEDSSI& *Saturated Structured Illumination &icroscopy5 SPE&/ 0(1(0r"un- (tate 1epleti"n) Temporal Resolution of STED Spatial resolution 6 field of 4ie7 2 scannin% rate 8urrent implementations allo7 (9ms lon% 4ideo-rate*+:fps/ recordin%s 7ith minimal resol4a$le distance of ';)nm and 9