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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Structured illumination in total internal reflection fluorescence microscopy using a spatial light modulator.
TL;DR: A versatile setup for standing-wave illumination in total internal reflection fluorescence microscopy with adjustable diffraction grating written on a phase-only spatial light modulator achieves 91 nm lateral resolution for green emission.
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Statistical analysis of molecule colocalization in bioimaging
TL;DR: It is shown on synthetic and biological images that object-based methods are more robust statistically than pixel‐based methods, and allow moreover to quantify accurately the number of colocalized molecules.
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Fluorescence Microscopy: A Concise Guide to Current Imaging Methods
Christian A. Combs,Hari Shroff +1 more
TL;DR: The field of fluorescence microscopy is rapidly growing and offers ever more imaging capabilities for biologists as discussed by the authors, and many new technologies and techniques have been developed that allow for combinations of deeper, faster, and higher resolution imaging.
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Fluorescence nanoscopy by polarization modulation and polarization angle narrowing.
Nour Hafi,Matthias Grunwald,Laura S van den Heuvel,Timo Aspelmeier,Jian-Hua Chen,Marta Zagrebelsky,Ole M. Schütte,Claudia Steinem,Martin Korte,Axel Munk,Peter Walla +10 more
TL;DR: Measurement of the average orientation of fluorescent dyes attached to rigid sample structures mapped to regularly defined image nanoareas can provide subdiffraction resolution and is applied to standard wide-field microscopy with camera detection and to two-photon scanning microscopy, imaging the fine structural details of neuronal spines.
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Microscopy beyond the diffraction limit using actively controlled single molecules
TL;DR: The addition of on/off control of molecular emission to maintain concentrations at very low levels in each imaging frame combined with sequential imaging of sparse subsets has enabled the reconstruction of images with resolution far below the optical diffraction limit.
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.
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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.
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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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