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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Journal ArticleDOI
Optogenetic Control of Synaptic Composition and Function.
Brooke L. Sinnen,Aaron B. Bowen,Jeffrey S. Forte,Brian G. Hiester,Kevin C. Crosby,Emily S. Gibson,Mark L. Dell'Acqua,Matthew J. Kennedy +7 more
TL;DR: An approach that uses light to tune the abundance of specific molecules in the postsynaptic density to support a model where adding AMPA receptors is sufficient to activate synapses that had few receptors to begin with, but that additional remodeling events are required to strengthen established synapses.
Journal ArticleDOI
Rational Design of Photoconvertible and Biphotochromic Fluorescent Proteins for Advanced Microscopy Applications
Virgile Adam,Benjamien Moeyaert,Charlotte C. David,Hideaki Mizuno,Mickaël Lelimousin,Peter Dedecker,Ryoko Ando,Atsushi Miyawaki,Jan Michiels,Yves Engelborghs,Johan Hofkens +10 more
TL;DR: NijiFP is presented, a promising new fluorescent protein with photoconvertible and biphotochromic properties that make it ideal for advanced fluorescence-based imaging applications and a new set of such enhanced optical highlighters derived from mEosFP and Dendra2.
Journal ArticleDOI
Wide-field multispectral super-resolution imaging using spin-dependent fluorescence in nanodiamonds.
Edward H. Chen,Ophir Gaathon,Ophir Gaathon,Matthew E. Trusheim,Matthew E. Trusheim,Dirk Englund,Dirk Englund +6 more
TL;DR: By modulating the fluorescence brightness of nitrogen-vacancy defect centers in nanodiamonds, a `deterministic emitter switch microscopy' (DESM) technique is demonstrated that enables super-resolution imaging with localization down to 47 nm across a 4×4μm2 area.
Journal ArticleDOI
Dual color localization microscopy of cellular nanostructures
Manuel Gunkel,Fabian Erdel,Karsten Rippe,Paul Lemmer,Rainer Kaufmann,Christoph Hörmann,Roman Amberger,Christoph Cremer +7 more
TL;DR: The capabilities of 2CLM for studying the spatial organization of the genome in the mammalian cell nucleus are demonstrated for the relative distributions of two chromosomal proteins labeled with autofluorescent GFP and mRFP1 domains.
Journal ArticleDOI
Single-particle tracking methods for the study of membrane receptors dynamics.
TL;DR: This work focuses on a qualitative description of the implementation of SPT, from molecule labelling to acquisition, data treatment and analysis of protein diffusion properties, and constraints, limitations and future developments are discussed.
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.
Journal ArticleDOI
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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