Far-Field Optical Nanoscopy by Philip Tinnefeld, Christian Eggeling, Stefan W. Hell

By Philip Tinnefeld, Christian Eggeling, Stefan W. Hell

This e-book describes advancements within the box of super-resolution fluorescence microscopy or nanoscopy. In eleven chapters, distinct scientists and leaders of their respective fields describe assorted nanoscopy ways, numerous labeling applied sciences, and urban purposes. the themes lined comprise the rules and purposes of the preferred nanoscopy concepts STED and (f)PALM/STORM, besides advances caused via fluorescent proteins and natural dyes optimized for fluorescence nanoscopy. in addition, the photophysics of fluorescent labels is addressed, in particular for making improvements to their photoswitching functions. vital purposes also are mentioned, reminiscent of the monitoring and counting of molecules to figure out appearing forces in cells, and quantitative mobile imaging, respectively, in addition to the mapping of chemical response facilities on the nano-scale.

The 2014 Chemistry Nobel Prize® was once provided for the ground-breaking developments of super-resolved fluorescence microscopy. during this publication, which used to be co-edited via one of many prize winners, readers will locate the newest advancements during this field.

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Opt Express 18(2):1302–1309 69. Bianchini P, Diaspro A (2012) Fast scanning STED and two-photon fluorescence excitation microscopy with continuous wave beam. J Microsc 245(3):225–228 70. Honigmann A, Eggeling C, Schulze M, Lepert A (2012) Super-resolution STED microscopy advances with yellow CW OPSL. Laser Focus World 48(1):75–79 71. Mueller V, Eggeling C, Karlsson H, von Gegerfelt D (2012) CW DPSS lasers make STED microscopy more practical. Biophotonics 19(5):30–32 72. Honigmann A, Mueller V, Fernando UP, Eggeling C, Sperling J (2013) Simplifying STED microscopy of photostable red-emitting labels.

ACS Nano. 1021/nn502856t 149. Vicidomini G, Ta H, Han KY, Moneron G, Honigmann A, Hell SW, Eggeling C (2014) Spatio-temporal heterogeneity of lipid membrane dynamics revealed by gated STED-FCS, in preparation 150. Hedde PN, Dorlich RM, Blomley R, Gradl D, Oppong E, Cato ACB, Nienhaus GU (2013) Stimulated emission depletion-based raster image correlation spectroscopy reveals biomolecular dynamics in live cells. Nat Commun 4:2093 151. Bianchini P, Cardarelli F, Luca D, Diaspro A, Bizzarri R (2014) Nanoscale protein diffusion by STED-based pair correlation analysis.

It is also worth noting that the localization density is not only limited by the label density but also by the on-off duty cycle of the photoswitchable probes, namely, the fraction of time each probe spends in the on state [40]. Because multiple probes emitting at the same time within a diffraction-limited area generate overlapping images that reduce the localization precision, fluorophores with a duty cycle of 1/N typically allow less than N molecules to be localized within a diffraction-limited area.

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