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
Reads0
Chats0
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
Citations
More filters
Journal ArticleDOI
Optical Super-Resolution Imaging of Surface Reactions.
Tao Chen,Bin Dong,Kuangcai Chen,Fei Zhao,Xiaodong Cheng,Changbei Ma,Seungah Lee,Peng Zhang,Seong Ho Kang,Ji Won Ha,Weilin Xu,Ning Fang +11 more
TL;DR: An overview of recent advances in super-resolution chemical imaging of surface reactions is provided, including chemical kinetics and reaction dynamics; active-site distributions on single-particle surfaces; and size-, shape-, and facet-dependent catalytic activities of individual nanocatalysts.
Journal ArticleDOI
Functional organization of cytoplasmic inclusion bodies in cells infected by respiratory syncytial virus
Vincent Rincheval,Mickaël Lelek,Elyanne Gault,Camille Bouillier,Delphine Sitterlin,Sabine Blouquit-Laye,Marie Galloux,Christophe Zimmer,Jean-François Eléouët,Marie-Anne Rameix-Welti +9 more
TL;DR: Respiratory syncytial virus induces formation of inclusion bodies (IBs) sheltering viral RNA synthesis that accumulate newly synthetized viral mRNA and the viral M2-1 protein but exclude viral genomic RNA and RNA polymerase complexes.
Journal ArticleDOI
Multiplexed 3D super-resolution imaging of whole cells using spinning disk confocal microscopy and DNA-PAINT
Florian Schueder,Florian Schueder,Juanita Lara-Gutiérrez,Juanita Lara-Gutiérrez,Brian J. Beliveau,Brian J. Beliveau,Sinem K. Saka,Sinem K. Saka,Hiroshi Sasaki,Hiroshi Sasaki,Johannes B. Woehrstein,Johannes B. Woehrstein,Maximilian T. Strauss,Maximilian T. Strauss,Heinrich Grabmayr,Heinrich Grabmayr,Peng Yin,Peng Yin,Ralf Jungmann,Ralf Jungmann +19 more
TL;DR: 3D DNA points accumulation for imaging in nanoscale topography (DNA-PAINT) with spinning disk confocal (SDC) hardware is combined to overcome this limitation and enable DNA-based super-resolution microscopy in whole cells using standard instrumentation.
Journal ArticleDOI
Parallel ptychographic reconstruction.
TL;DR: A parallel method for real-time ptychographic phase retrieval that uses a hybrid parallel strategy to divide the computation between multiple graphics processing units (GPUs) and then employs novel techniques to merge sub-datasets into a single complex phase and amplitude image.
Journal ArticleDOI
Nanoscale segregation of actin nucleation and elongation factors determines dendritic spine protrusion
Anaël Chazeau,Amine Mehidi,Deepak T. Nair,Jérémie Gautier,Cécile Leduc,Ingrid Chamma,Frieda Kage,Adel Kechkar,Olivier Thoumine,Klemens Rottner,Daniel Choquet,Alexis Gautreau,Jean-Baptiste Sibarita,Grégory Giannone +13 more
TL;DR: The nanoscale organization and dynamics of branched F‐actin regulators in spines might be reorganized during spine morphological remodeling often associated with synaptic plasticity.
References
More filters
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.
Related Papers (5)
Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM).
Ultra-High Resolution Imaging by Fluorescence Photoactivation Localization Microscopy
Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy
Stefan W. Hell,Jan Wichmann +1 more