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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Photoswitchable Fluorescent Proteins: Ten Years of Colorful Chemistry and Exciting Applications
Xin X. Zhou,Michael Z. Lin +1 more
TL;DR: Structural studies reveal diversity in the details of photoswitching mechanisms, including different effects of protonation, chromophore planarity, and pocket flexibility in RSFPs.
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Correlative Super-Resolution Microscopy: New Dimensions and New Opportunities.
TL;DR: It is demonstrated how a correlative approach adds new dimensions of information and provides new opportunities in the fast-growing field of SRM.
Journal ArticleDOI
Fluorescence and super-resolution standards based on DNA origami
Jürgen J. Schmied,Andreas Gietl,Phil Holzmeister,Carsten Forthmann,Christian Steinhauer,Thorben Dammeyer,Philip Tinnefeld +6 more
TL;DR: This work presents molecular rulers based on self-assembled DNA origami structures as a general and highly versatile platform for fluorescence and super-resolution standards that are adapted to the needs of the specific microscopy technique, including stimulated emission depletion (STED), localization-based super- resolution and diffractionlimited microscopy.
Journal ArticleDOI
3D single-molecule super-resolution microscopy with a tilted light sheet.
Anna-Karin Gustavsson,Petar N. Petrov,Maurice Y. Lee,Yoav Shechtman,Yoav Shechtman,W. E. Moerner +5 more
TL;DR: The authors validate TILT3D for 3D super-resolution imaging in mammalian cells by imaging mitochondria and the full nuclear lamina using the double-helix PSF for single-molecule detection and the recently developed tetrapod PSFs for fiducial bead tracking and live axial drift correction.
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Visualization of localization microscopy data.
TL;DR: Two additional visualization algorithms are presented, an adaptive histogram method based on quad-trees and a Delaunay triangulation based visualization of point data that address some of the deficiencies of existing methods and are designed to suppress erroneous detail in poorly sampled image areas but avoid loss of resolution in well-sampled regions.
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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