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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Ultrasensitive Label-Free Nanosensing and High-Speed Tracking of Single Proteins
TL;DR: In this article, an all-optical method capable of detecting individual nanoparticles as small as 15 kDa proteins that is equivalent to half a GFP was proposed. But the method requires high-speed image acquisition exceeding several hundreds of frames-per-second.
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Photoactivation mechanism of PAmCherry based on crystal structures of the protein in the dark and fluorescent states.
Fedor V. Subach,Vladimir N. Malashkevich,Wendy D. Zencheck,Hui Xiao,Grigory S. Filonov,Steven C. Almo,Vladislav V. Verkhusha +6 more
TL;DR: Data indicate that PAmCherry1 in the dark state possesses the chromophore N-[(E)-(5-hydroxy-1H-imidazol-2-yl)methylidene]acetamide, which, to the authors' knowledge, has not been previously observed in PAFPs.
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Encoding and decoding spatio-temporal information for super-resolution microscopy
Luca Lanzano,Iván Coto Hernández,Marco Castello,Enrico Gratton,Alberto Diaspro,Giuseppe Vicidomini +5 more
TL;DR: The proposed method provides nanoscale imaging of subcellular structures, opening new routes in super-resolution microscopy based on the encoding/decoding of spatial information through manipulation of molecular dynamics.
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Absorption spectroscopy and imaging from the visible through mid-infrared with 20 nm resolution.
TL;DR: This work has reported the highest resolution reported for PTIR, as determined by comparing height and PTIR images, and its first extension to near-IR and visible wavelengths.
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Transfection of living HeLa cells with fluorescent poly-cytosine encapsulated Ag nanoclusters.
TL;DR: Bright near-infrared fluorescence was observed from inside the transfected HeLa cells, when exciting with 633 nm excitation, opening up the possibility for the use of these C(24):Ag(n) clusters for biological labelling and imaging of living cells and for monitoring theTransfection process with limited harm to the living cells.
References
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Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy.
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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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