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
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
Citations
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Ultra-High Resolution Imaging by Fluorescence Photoactivation Localization Microscopy
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Far-Field Optical Nanoscopy
TL;DR: Initial applications indicate that emergent far-field optical nanoscopy will have a strong impact in the life sciences and in other areas benefiting from nanoscale visualization.
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Three-Dimensional Super-Resolution Imaging by Stochastic Optical Reconstruction Microscopy
TL;DR: 3D stochastic optical reconstruction microscopy (STORM) is demonstrated by using optical astigmatism to determine both axial and lateral positions of individual fluorophores with nanometer accuracy, allowing the 3D morphology of nanoscopic cellular structures to be resolved.
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Antennas for light
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TL;DR: Optical antennas are devices that convert freely propagating optical radiation into localized energy, and vice versa as mentioned in this paper, and hold promise for enhancing the performance and efficiency of photodetection, light emission and sensing.
References
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Use of caged fluorochromes to track macromolecular movement in living cells.
TL;DR: The use of photoactivatable fluorochromes to track the intracellular movement of both proteins and nucleic acids and to track cell lineages is described.
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Far-field fluorescence microscopy beyond the diffraction limit
TL;DR: In this paper, a technique was developed to obtain three-dimensional structural information on a length scale well below the Rayleigh length with conventional far-field optics by spectrally selecting a single molecule with high-resolution laser spectroscopy and using a CCD camera to register the spatial distribution of the emitted photons in three dimensions.
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Far-field fluorescence microscopy beyond the diffraction limit
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Excitation strategies for optical lattice microscopy
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