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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Super-resolution fight club: assessment of 2D and 3D single-molecule localization microscopy software.
Daniel Sage,Thanh-an Pham,Hazen P. Babcock,Tomas Lukes,Tomas Lukes,Thomas Pengo,Jerry Chao,Ramraj Velmurugan,Alex Herbert,Anurag Agrawal,Silvia Colabrese,Silvia Colabrese,Ann P. Wheeler,Anna Archetti,Bernd Rieger,Raimund J. Ober,Raimund J. Ober,Guy M. Hagen,Jean-Baptiste Sibarita,Jean-Baptiste Sibarita,Jonas Ries,Ricardo Henriques,Michael Unser,Seamus Holden +23 more
TL;DR: This study reports results from the second community-wide single-molecule localization microscopy software challenge, which tested over 30 software packages on realistic simulated data for multiple popular 3D image acquisition modes, as well as 2D localization microscopes.
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3‐D PSF fitting for fluorescence microscopy: implementation and localization application
TL;DR: A least‐squares point spread function fitting framework that utilizes the Gibson and Lanni model and a computationally efficient way for evaluating its derivative functions is introduced and demonstrated with algorithms for particle localization and defocus estimation.
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Super-resolution microscopy at a glance.
TL;DR: Advances in microscopy and cell biology are intimately intertwined, with new visualization possibilities often leading to dramatic leaps in the understanding of how cells function.
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Superpenetration optical microscopy by iterative multiphoton adaptive compensation technique
TL;DR: It is shown that high quality three-dimensional imaging can be realized at depths beyond the reach of conventional multiphoton microscopy and adaptive optics methods, albeit over restricted distances for a given correction.
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Super-Resolution Microscopy: From Single Molecules to Supramolecular Assemblies
TL;DR: This review surveys the application of SRM in elucidating the structure of macromolecules in the native cellular environment and discusses both the novel information that can be generated through SRM as well as the experimental considerations to examine while conducting such studies.
References
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疟原虫var基因转换速率变化导致抗原变异[英]/Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A
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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