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
Functional Nanoscale Organization of Signaling Molecules Downstream of the T Cell Antigen Receptor
Eilon Sherman,Valarie A. Barr,Suliana Manley,George H. Patterson,Lakshmi Balagopalan,Itoro Akpan,Carole K. Regan,Robert K. Merrill,Connie L. Sommers,Jennifer Lippincott-Schwartz,Lawrence E. Samelson +10 more
TL;DR: This work used single and two-color photoactivated localization microscopy to study complexes downstream of the T cell antigen receptor (TCR) in single-molecule detail at the plasma membrane of intact T cells to extend the understanding of the mechanism of T cell activation and the formation and organization of TCR-mediated signaling complexes.
Journal ArticleDOI
Tracking single molecules at work in living cells
TL;DR: SMT methods are reviewed, the recent results obtained by SMT are summarized, related superresolution microscopy data is summarized, and special concerns when SMT applications are shifted from the in vitro paradigms to living cells are described.
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Genetically encoded biosensors based on engineered fluorescent proteins
TL;DR: This tutorial review provides an overview of the progress made in the development of fluorescent protein-based biosensors to date.
Journal ArticleDOI
Fourier ring correlation as a resolution criterion for super-resolution microscopy.
TL;DR: It is shown that Fourier ring correlation provides an easy-to-use, laboratory consistent standard for measuring the resolution of SRM images, and is provided a freely available software tool that combines resolution measurement with image reconstruction.
Journal ArticleDOI
Optochemical Control of Biological Processes in Cells and Animals.
TL;DR: Recent developments of optochemical tools, including small molecules, peptides, proteins, and nucleic acids that can be irreversibly or reversibly controlled through light irradiation, are discussed, with a focus on applications in cells and animals.
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.
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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.
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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