Resources Cryo & 3D Electron Tomography

Investigations & Applications

3D Electron Microscopy Research Projects

Summaries of in-house research projects using electron tomography and cryo electron microscopy to study the invaginations in the mitochondrial inner membrane, functional states of the ryanodine receptor (RyR), the interaction between the ribosome and its ligands and kinetochore modulation of microtubule dynamics. An extensive list of publications is included in the site.

From: , Wadsworth Center
Wadsworth Center
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Three-dimensional electron crystallography of protein microcrystals

Using a new protocal electron diffraction data from protein crystals was collected at 2.9 Å resolution using electron diffraction in a cryo electron microscope and result in. The protocol used an electron diffraction collected at resolutions of 1.7 Å. A full tilt series of 90 diffraction patterns was collected from three crystals. The technique is named ‘MicroED’.

From: , HHMI Janelia Farm Research Campus
HHMI Janelia Farm Research Campus
Authors: Dan Shi, Brent L Nannenga, Matthew GIadanza, Tamir Gonen
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Web Portals

3D-EM Web Site

The site includes information on the 3D EM mailing list, laboraotries and events.

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Introductions and Briefs

Cryo-electron tomography and Single-particle cryo-EM

The techniques development section contains summaries of the contributions of scientist at the Wadworth Center to the development of three-dimensional reconstruction of single particles and cryo-electron tomography techniques. A list of references is included.

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Advances in high-resolution imaging – techniques for three-dimensional imaging of cellular structures

This 2012 commentary provides an excellent overview of microscopic techniques used in the 3D imaging of cellular structures. Advances in electron microscopy tomography and fluorescence super-resolution techniques including STED, Single-molecule-localization-based super-resolution, Plane illumination microscopy, and Structured illumination microscopy are reviewed. There is an insightful comparison of super resolution techniques to confocal microscopy in terms of difficulty of preparation, processing and live cell.

From: Super-Resolution Technology Core, University of New Mexico
Super-Resolution Technology Core, University of New Mexico
Authors: Diane S. Lidke and Keith A. Lidke
Citation: June 1, 2012 J Cell Sci 125, 2571-2580., doi: 10.1242/​jcs.090027
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Technical Overviews

Imaging the building blocks of viruses

New microscopy techniques are illuminating viral structures and protein interaction. Surface plasmon resonance based on the optical excitation of surface plasmons is being used to detect high-resolution, label-free imaging of single viruses. Cryo electron microscopy, pushed to a resolution of 4Å can image amino acid side chains in proteins. X-ray techniques now are now able to to provide data on virus that are pieced together to form a image of the structure.

Authors: Suzanne Winter
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Electron Cryotomography

Peer reviewed article summarizing using cryoelectron microscopy to reconstruct 3-D structures of macromolcules, virus and small whole cells. Includes are details on the insstrumentation, preparation, limitations, and data collection.

From: , California Institute of Technology
California Institute of Technology
Authors: Gavin E. Murphy and Grant J. Jensen
Citation: BioTechniques, Vol. 43 ı No. 4
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Direct detection pays off for electron cryo-microscopy

This online journal article described improvements in image-processing techniques and electron detectors that decrease the number of images needed for single particle cryo-electron microscopy images.

From: , Brandeis University
Brandeis University
Authors: Nikolaus Grigorieff
Citation: http://dx.doi.org/10.7554/eLife.00573
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A national facility for biological cryo-electron microscopy

An excellent introduction to the techniques of cryo-electron microscopy is included in this paper on the electron Bio-Imaging Centre at Diamond Light Source. Topics reviewed are single particle analysis, electron cryo-tomography, correlative microscopy, and electron crystallography.

From: electron Bio-Imaging Centre eBIC, Diamond Light Source
electron Bio-Imaging Centre eBIC, Diamond Light Source
Authors: Helen R. Saibil,a* Kay Grünewaldb and David I. Stuartb,c
Citation: Biological Crystallography, Volume 71, January 2015, 127-135 doi:10.1107/S1399004714025280
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Software

Software Tools For Molecular Microscopy

The Wiki Open Content document is a comprehensive list of the software developed for cryo-electron microscopy and molecular microscopy. Software is loosely categorized by functionality. Website, contact, description, cost, and references are included for each tool. The page is well supported by the scientific community.

From: National Resource for Automated Molecular Microscopy, New York Structural Biology Center
National Resource for Automated Molecular Microscopy, New York Structural Biology Center
Authors: Voss NR, Potter CS, Smith R, Carragher B.
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