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3D Microscopy

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  • Measuring the 3D STED-PSF with a new Type of Fluorescent Beads

    A new type of fluorescent bead is presented by GATTAquant. These beads, called GATTA-Beads, are characterized by a small diameter (23 nm), high intensity and size uniformity. In combination with state-of the-art STED microscopes such as the Leica TCS SP8 STED 3X and high-end image restoration methods available in the Huygens Software, it is shown that these new beads can be used for accurate STED PSF characterization in 3D. Furthermore, it is shown that the measured 3D STED-PSF can be used to improve image restoration quality in combination with STED deconvolution methods available in the Huygens Software.
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  • 4Pi-RESOLFT Nanoscopy

    Here we apply the 4Pi scheme to RESOLFT nanoscopy using two-photon absorption for the on-switching of fluorescent proteins. We show that in this combination, the lobes are so low that low-light level, 3D nanoscale imaging of living cells becomes possible. Our method thus offers robust access to densely packed, axially extended cellular regions that have been notoriously difficult to super-resolve. Our approach also entails a fluorescence read-out scheme that translates molecular sensitivity to local off-switching rates into improved signal-to-noise ratio and resolution.
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  • Video Interviews with Kees Jalink

    Kees Jalink's group at the Netherlands Cancer Institute in Amsterdam, The Netherlands, explores signal transduction pathways and cell adhesion processes in cancer cells. In his eyes especially the new three-dimensional nanoscopic view of the relevant structure of interest is an essential feature to get the full picture.
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  • Abstracts of the 4th European Super-Resolution User-Club Meeting

    The 4th Super-Resolution User Club Meeting was held in collaboration with Christian Eggeling and the Weatherall Institute of Molecular Medicine in Oxford, UK. Here we present the abstracts of the talks and interviews with participants.
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  • Super-resolution Microscopy and the Third Dimension

    Optical imaging devices have a finite depth of field and diffraction limited resolution. The depth of field problem was tackled first with confocal microscopes, diffraction unlimited resolution is available since a few years with super-resolution microscopes. Super-resolution microscopes with a solved depth of field problem are now available.
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  • Three-Dimensional Super-Resolution GSDIM Microscopy

    With the new 3D GSDIM technique structures like the Golgi and the microtubular network are resolved not only laterally, but also in a third dimension. The principle is based on the use of optical astigmatism to determine the accurate lateral and axial position of individual fluorochromes.
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  • 3D Localization Microscopy With Ground State Depletion (GSD)

    With the latest development of a GSD 3D super-resolution platform, it is now possible to achieve a lateral resolution of down to 20 nm and an axial resolution of 70 nm. The technology is based on an astigmatism approach using a manipulated PSF to localize the molecule in z. This following tutorial describes the basic principles of the 3D GSD technology.
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  • New Opportunities for 3D Technology in Medicine

    Until now, physicians have largely been skeptical of the advantages of 3D technology. But this may be about to change: the findings of a new study show that even experienced surgeons stand to benefit from the third dimension.
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  • A New FRAP/FRAPa Method for Three-Dimensional Diffusion Measurements Based on Multiphoton Excitation Microscopy

    Quantitative measurement method based on FRAP and FRAPa using multiphoton microscopy. We present a new convenient method for quantitative three-dimensionally resolved diffusion measurements based on the photobleaching (FRAP) or photoactivation (FRAPa) of a disk-shaped area by the scanning laser beam of a multiphoton microscope. Contrary to previously reported spot-photobleaching protocols, this method has the advantage of full scalability of the size of the photobleached area and thus the range of diffusion coefficients, which can be measured conveniently.
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