2015
DOI: 10.1017/s0033583514000146
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Lens-based fluorescence nanoscopy

Abstract: The majority of studies of the living cell rely on capturing images using fluorescence microscopy. Unfortunately, for centuries, diffraction of light was limiting the spatial resolution in the optical microscope: structural and molecular details much finer than about half the wavelength of visible light (~200 nm) could not be visualized, imposing significant limitations on this otherwise so promising method. The surpassing of this resolution limit in far-field microscopy is currently one of the most momentous … Show more

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Cited by 140 publications
(143 citation statements)
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References 435 publications
(1,084 reference statements)
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“…We incorporated the Chol analogs into an SLB, as before, and measured their average transit times ( D ) as we increased the power ( P STED ) of the STED laser. From theory, the diameter ( d ) of the observation spot decreases with increasing P STED ( d ‫ف‬ C/ √ P STED , with C depending on, among other parameters, the photophysical properties of the fl uorescent label) ( 79 ). Figure 4E, F plots the dependencies [ d ( P STED )] obtained for the different Chol analogs.…”
Section: Fcs On Chol Analogsmentioning
confidence: 99%
See 1 more Smart Citation
“…We incorporated the Chol analogs into an SLB, as before, and measured their average transit times ( D ) as we increased the power ( P STED ) of the STED laser. From theory, the diameter ( d ) of the observation spot decreases with increasing P STED ( d ‫ف‬ C/ √ P STED , with C depending on, among other parameters, the photophysical properties of the fl uorescent label) ( 79 ). Figure 4E, F plots the dependencies [ d ( P STED )] obtained for the different Chol analogs.…”
Section: Fcs On Chol Analogsmentioning
confidence: 99%
“…On one hand, reduced observation spot sizes yield a smaller amount of total photons from a traversing single molecule. On the other hand, the additional STED laser irradiation might induce additive photobleaching ( 30,79 ). We checked the performance of those Chol analogs that we had already tested in the conventional FCS recordings for their usefulness for STED-FCS.…”
Section: Fcs On Chol Analogsmentioning
confidence: 99%
“…In the so-called targeted approach [17], light patterns are applied to the sample that confine the detection of fluorophores to regions more confined than with conventional microscopy. In conventional raster scanning confocal far-field fluorescence microscopy, a tightly focused laser spot (Airy spot) is raster-scanned across the sample, pixel by pixel, to excite fluorophores, and the photons emitted from each pixel are collected to reconstruct the final image.…”
Section: A Brief Survey Of Super-resolution Fluorescence Imaging Methodsmentioning
confidence: 99%
“…Nevertheless, all of these techniques rely on optical detection and hence are limited to detecting the dynamic behavior of optical markers attached to the molecules. Although super-resolution fluorescence microscopy breaking the diffraction limit of light has been recently added to the growing list of tools (Eggeling et al 2015), this limitation cannot be eliminated even by this new microscopy.…”
Section: Introductionmentioning
confidence: 99%