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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Lattice Light Sheet Microscopy: Imaging Molecules to Embryos at High Spatiotemporal Resolution
Bi-Chang Chen,Wesley R. Legant,Kai Wang,Lin Shao,Daniel E. Milkie,Michael W. Davidson,Chris Janetopoulos,Xufeng S. Wu,John A. Hammer,Zhe Liu,Brian P. English,Yuko Mimori-Kiyosue,Daniel P. Romero,Alex T. Ritter,Alex T. Ritter,Jennifer Lippincott-Schwartz,Lillian K. Fritz-Laylin,R. Dyche Mullins,Diana M. Mitchell,Joshua N. Bembenek,Anne-Cécile Reymann,Ralph Böhme,Stephan W. Grill,Jennifer T. Wang,Geraldine Seydoux,U. Serdar Tulu,Daniel P. Kiehart,Eric Betzig +27 more
TL;DR: A new microscope using ultrathin light sheets derived from two-dimensional optical lattices is developed, demonstrating the performance advantages of lattice light-sheet microscopy compared with previous techniques and highlighted phenomena that, when seen at increased spatiotemporal detail, may hint at previously unknown biological mechanisms.
Journal ArticleDOI
Spatially resolved, highly multiplexed RNA profiling in single cells
Kok Hao Chen,Alistair N. Boettiger,Jeffrey R. Moffitt,Siyuan Wang,Xiaowei Zhuang,Xiaowei Zhuang +5 more
TL;DR: This report reports multiplexed error-robust FISH (MERFISH), a single-molecule imaging method that allows thousands of RNA species to be imaged in single cells by using combinatorial FISH labeling with encoding schemes capable of detecting and/or correcting errors.
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Super-Resolution Fluorescence Microscopy
TL;DR: It is anticipated that super-resolution fluorescence microscopy will become a widely used tool for cell and tissue imaging to provide previously unobserved details of biological structures and processes.
Journal ArticleDOI
Multicolor Super-Resolution Imaging with Photo-Switchable Fluorescent Probes
TL;DR: A family of photo-switchable fluorescent probes is introduced and multicolor stochastic optical reconstruction microscopy (STORM) is demonstrated, to facilitate direct visualization of molecular interactions at the nanometer scale.
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Spiropyran-based dynamic materials
TL;DR: This review discusses the synthesis, switching conditions, and use of dynamic materials in which spiropyran has been attached to the surfaces of polymers, biomacromolecules, inorganic nanoparticles, as well as solid surfaces.
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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