Foundations of STED microscopy

Abstract

This chapter presents the foundations of STED microscopy with a comparison to its generalization Resolft microscopy and to stochastic imaging methods (PALM, STORM, FPALM, and alike). The first section reviews the advantages of optical microscopy, explains the diffraction limit, and shows how the classical resolution limit was finally broken. It also reviews some of the achievements in super-resolution imaging. The second section explains in depth the principle of STED microscopy and highlights some special STED modalities like the use of continuous wave lasers, time-gating, fast imaging with up to 200 frames per second, and combination with fluorescence correlation spectroscopy. The third section treats some aspects of resolution in the presence of noise, especially in the scope of high-resolution imaging. © 2014 Springer Science+Business Media, LLC.