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
Counting the Number of Proteins Coupled to Single Nanoparticles
Didier Casanova,Domitille Giaume,Mélanie Moreau,Jean-Louis Martin,Thierry Gacoin,Jean-Pierre Boilot,Antigoni Alexandrou +6 more
TL;DR: The accurate quantification of the biomolecule−particle coupling opens up the possibility of selecting finely controlled conjugates and the distribution of the protein−nanoparticle ratio was precisely measured.
Journal ArticleDOI
Super-resolution spectroscopic microscopy via photon localization.
Biqin Dong,Luay M. Almassalha,Ben E. Urban,The-Quyen Nguyen,Satya Khuon,Teng-Leong Chew,Vadim Backman,Cheng Sun,Hao Zhang +8 more
TL;DR: Using spectroscopic photon localization microscopy, simultaneous multi-colour super-resolution imaging of microtubules and mitochondria in COS-7 cells is demonstrated and it is shown that background autofluorescence can be identified through its distinct emission spectra.
Book ChapterDOI
Functional nuclear architecture studied by microscopy: present and future.
TL;DR: In this article, major contributions of light and electron microscopic approaches to the present understanding of functional nuclear architecture are described. But the large gap of knowledge, which must still be bridged from the molecular level to the level of higher order structure, is emphasized by differences of currently discussed models of nuclear architecture.
Journal ArticleDOI
PEGylation and cell imaging applications of AIE based fluorescent organic nanoparticles via ring-opening reaction
Xiaoyong Zhang,Xiaoyong Zhang,Xiqi Zhang,Bin Yang,Junfeng Hui,Meiying Liu,Wanyun Liu,Yiwang Chen,Yen Wei +8 more
TL;DR: PEGylation of aggregation induced emission based fluorescent organic nanoparticles via one pot ring-opening polymerization and condensation reaction was developed and exhibited high water dispersibility, strong fluorescence, uniform morphology and more important excellent biocompatibility, implying their high potential for various biomedical applications.
Journal ArticleDOI
Towards correlative super-resolution fluorescence and electron cryo-microscopy
TL;DR: A major obstacle of cryo‐CLEM currently hindering many biological applications is the large resolution gap betweenCryo‐FM (typically in the range of ∼400 nm) and cryo-EM (single nanometre to the Ångstrom range).
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.
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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