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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Two-color nanoscopy of three-dimensional volumes by 4Pi detection of stochastically switched fluorophores
Daniel Aquino,Andreas Schönle,Claudia Geisler,Claas von Middendorff,Christian A. Wurm,Yosuke Okamura,Thorsten Lang,Stefan W. Hell,Alexander Egner +8 more
TL;DR: Three-dimensional super-resolution imaging of stochastically switched fluorophores distributed across whole cells and of other transparent materials is demonstrated, offering a combination of multicolor recording, nanoscale resolution and extended axial depth.
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Accuracy of the Gaussian Point Spread Function model in 2D localization microscopy
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TL;DR: The effect of emission dipole orientation in conjunction with optical aberrations on the localization accuracy of position estimators based on a gaussian model PSF is studied.
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Widely accessible method for superresolution fluorescence imaging of living systems
TL;DR: This method, which uses off-the-shelf equipment, genetically encodable labels, and simple and rapid data acquisition, is capable of providing two- to threefold-enhanced spatial resolution, significant background rejection, markedly improved contrast, and favorable temporal resolution in living cells.
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Polymerizable aggregation-induced emission dye-based fluorescent nanoparticles for cell imaging applications
Xiaoyong Zhang,Xiaoyong Zhang,Xiqi Zhang,Bin Yang,Meiying Liu,Wanyun Liu,Yiwang Chen,Yen Wei +7 more
TL;DR: A polymerizable aggregation-induced emission (AIE) dye (named PhE) was facilely incorporated into polymer nanoparticles through reversible addition fragmentation chain transfer polymerization and showed uniform size, high water dispersibility, strong fluorescence and excellent biocompatibility.
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Super-resolution Microscopy Approaches for Live Cell Imaging
TL;DR: This mini-review describes and compares the main far-field super-resolution approaches that allow studying endogenous or overexpressed proteins in live 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
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Journal ArticleDOI
Nonlinear structured-illumination microscopy: Wide-field fluorescence imaging with theoretically unlimited resolution
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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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