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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Single-Molecule Fluorescence Imaging in Living Cells
Tie Xia,Nan Li,Xiaohong Fang +2 more
TL;DR: The practical considerations in designing single-molecule fluorescence imaging in cells are discussed, including the choice of fluorescent probes, labeling methods, instrumentation, and imaging techniques, and what can be learned from such characterizations.
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DNA origami–based standards for quantitative fluorescence microscopy
Jürgen J. Schmied,Mario Raab,Carsten Forthmann,Enrico Pibiri,Bettina Wünsch,Thorben Dammeyer,Philip Tinnefeld +6 more
TL;DR: The protocol provides procedures for the design and realization of fluorescent marks on DNA origami structures, their production and purification, quality control, handling, immobilization, measurement and data analysis.
Journal ArticleDOI
Coordinate-based colocalization analysis of single-molecule localization microscopy data
Sebastian Malkusch,Ulrike Endesfelder,Justine Mondry,Márton Gelléri,Peter J. Verveer,Mike Heilemann +5 more
TL;DR: An algorithm for coordinate-based colocalization analysis which is suited for single-molecule super-resolution data is introduced and an experimental configuration for simultaneous dual-color imaging together with a robust approach to correct for optical aberrations with a few nanometers is presented.
Journal ArticleDOI
Lensless wide-field fluorescent imaging on a chip using compressive decoding of sparse objects
TL;DR: A compressive sampling based optimization algorithm is used to rapidly reconstruct the sparse distribution of fluorescent sources to achieve ~10 µm spatial resolution over the entire active region of the sensor-array over an imaging field-of-view of >8 cm2.
Journal ArticleDOI
Nanoscale protein architecture of the kidney glomerular basement membrane
Hani Suleiman,Lei Zhang,Robyn Roth,John E. Heuser,Jeffrey H. Miner,Andrey S. Shaw,Adish Dani +6 more
TL;DR: Sub-diffraction resolution stochastic optical reconstruction microscopy (STORM) is used to resolve the in situ molecular organization of proteins within the kidney glomerular basement membrane (GBM), an essential mediator ofglomerular ultrafiltration and provides the first nanoscopic glimpse into the organization of a complex ECM.
References
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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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