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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Nanoscopy of Living Brain Slices with Low Light Levels
Ilaria Testa,Nicolai T. Urban,Stefan Jakobs,Christian Eggeling,Katrin I. Willig,Stefan W. Hell +5 more
TL;DR: The superresolution fluorescence microscopy called RESOLFT enables comparatively fast and continuous imaging of sensitive, nanosized features in living brain tissue and enables the recording of spontaneous and stimulated changes of dendritic actin filaments and spine morphology occurring on time scales from seconds to hours.
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Parallelized STED fluorescence nanoscopy.
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Reversible photoswitching in fluorescent proteins: A mechanistic view
TL;DR: These proteins offer the widest range of practical applications, including reversible high‐density data bio‐storage, photochromic FRET, and super‐resolution microscopy by either point‐scanning, structured illumination, or single molecule‐based wide‐field approaches.
Journal ArticleDOI
Three-dimensional localization precision of the double-helix point spread function versus astigmatism and biplane
TL;DR: The theoretical localization precision for an unbiased estimator of the DH-PSF is compared to that for 3D localization by astigmatic and biplane imaging using Fisher information analysis including pixelation and varying levels of background.
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The role of molecular dipole orientation in single-molecule fluorescence microscopy and implications for super-resolution imaging.
TL;DR: A number of methods for measuring molecular orientation using fluorescence microscopy are reviewed, focusing on approaches that are most compatible with position estimation and single-molecule super-resolution imaging.
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.
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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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