Imaging intracellular fluorescent proteins at nanometer resolution.
Eric Betzig,George H. Patterson,Rachid Sougrat,O. Wolf Lindwasser,Scott G. Olenych,Juan S. Bonifacino,Michael W. Davidson,Jennifer Lippincott-Schwartz,Harald F. Hess +8 more
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TLDR
This work introduced a method for optically imaging intracellular proteins at nanometer spatial resolution and used this method to image specific target proteins in thin sections of lysosomes and mitochondria and in fixed whole cells to image retroviral protein Gag at the plasma membrane.Abstract:
We introduce a method for optically imaging intracellular proteins at nanometer spatial resolution. Numerous sparse subsets of photoactivatable fluorescent protein molecules were activated, localized (to approximately 2 to 25 nanometers), and then bleached. The aggregate position information from all subsets was then assembled into a superresolution image. We used this method--termed photoactivated localization microscopy--to image specific target proteins in thin sections of lysosomes and mitochondria; in fixed whole cells, we imaged vinculin at focal adhesions, actin within a lamellipodium, and the distribution of the retroviral protein Gag at the plasma membrane.read more
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Fluorescence Nanoscopy in Whole Cells by Asynchronous Localization of Photoswitching Emitters
Alexander Egner,Claudia Geisler,Claas von Middendorff,Hannes Bock,Dirk Wenzel,Rebecca Medda,Martin Andresen,Andre C. Stiel,Stefan Jakobs,Christian Eggeling,Andreas Schönle,Stefan W. Hell +11 more
TL;DR: N nanoscale resolution in far-field fluorescence microscopy is demonstrated using reversible photoswitching and localization of individual fluorophores at comparatively fast recording speeds and from the interior of intact cells by asynchronously recording the photon bursts of individual molecular switching cycles.
Journal ArticleDOI
Ultra-stable organic fluorophores for single-molecule research
Qinsi Zheng,Manuel F. Juette,Steffen Jockusch,Michael R. Wasserman,Zhou Zhou,Roger B. Altman,Scott C. Blanchard +6 more
TL;DR: Self-healing organic fluorophores, wherein the triplet state is intramolecularly quenched by a covalently attached protective agent, exhibit markedly improved photostabilities and are likely to impact the future of single-molecule research.
Journal ArticleDOI
Advances in Light Microscopy for Neuroscience
Brian A. Wilt,Laurie D. Burns,Eric Tatt Wei Ho,Kunal K. Ghosh,Eran A. Mukamel,Mark J. Schnitzer +5 more
TL;DR: Progress in Mouse genetic approaches permit mosaic and inducible fluorescence-labeling strategies, whereas intrinsic contrast mechanisms allow in vivo imaging of animals and humans without use of exogenous markers.
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Time-lapse two-color 3D imaging of live cells with doubled resolution using structured illumination
TL;DR: A previously undescribed SIM setup that is fast enough to record 3D two-color datasets of living whole cells and shows volume rates as high as 4 s in one color and 8.5 s in two colors over tens of time points is reported.
Journal ArticleDOI
Noninvasive Imaging beyond the Diffraction Limit of 3D Dynamics in Thickly Fluorescent Specimens
Liang Gao,Lin Shao,Christopher D. Higgins,John S. Poulton,Mark Peifer,Michael W. Davidson,Xufeng Wu,Bob Goldstein,Eric Betzig +8 more
TL;DR: It is reported that rapid three-dimensional dynamics can be studied beyond the diffraction limit in thick or densely fluorescent living specimens over many time points by combining ultrathin planar illumination produced by scanned Bessel beams with super-resolution structured illumination microscopy.
References
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TL;DR: PfPMP1)与感染红细胞、树突状组胞以及胎盘的单个或多个受体作用,在黏附及免疫逃避中起关键的作�ly.
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Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy.
TL;DR: Lateral resolution that exceeds the classical diffraction limit by a factor of two is achieved by using spatially structured illumination in a wide‐field fluorescence microscope with strikingly increased clarity compared to both conventional and confocal microscopes.
Journal ArticleDOI
Precise nanometer localization analysis for individual fluorescent probes
TL;DR: A localization algorithm motivated from least-squares fitting theory is constructed and tested both on image stacks of 30-nm fluorescent beads and on computer-generated images (Monte Carlo simulations), and results show good agreement with the derived precision equation.
Journal ArticleDOI
Nonlinear structured-illumination microscopy: Wide-field fluorescence imaging with theoretically unlimited resolution
TL;DR: Experimental results show that a 2D point resolution of <50 nm is possible on sufficiently bright and photostable samples, and a recently proposed method in which the nonlinearity arises from saturation of the excited state is experimentally demonstrated.
Journal ArticleDOI
Myosin V Walks Hand-Over-Hand: Single Fluorophore Imaging with 1.5-nm Localization
Ahmet Yildiz,Joseph N. Forkey,Sean A. McKinney,Taekjip Ha,Taekjip Ha,Yale E. Goldman,Paul R. Selvin,Paul R. Selvin +7 more
TL;DR: The results strongly support a hand-over-hand model of motility, not an inchworm model, which moves processively on actin.
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