Techniques Overview
Structural biology techniques address sample diversity. Critical structures and functions in biology occur across a wide range of distances (subnanometers to centimeters) and times (subpicoseconds to minutes). The DOE Biological and Environmental Research Program makes available a variety of structural biology techniques suited to investigating different sample lengths and experimental time scales. [See bottom of page for image credits.]
At these facilities, BER supports beamlines and resources to enable multiscale structural studies. New insights linking molecular properties to system-level functions can be achieved by integrating these different capabilities to make cross-scale connections and leveraging data from complementary techniques such as super-resolution optical and magnetic resonance imaging.
Cryo-Electron Microscopy and Tomography
Electrons enable sample imaging from nucleic acids to large biological assemblies frozen in their native states, at nanometer to atomic scales.
Resources offering this technique
Additional enabling capabilities
Hard X-Ray Tomography
A non-invasive full-field imaging technique used to measure the insides of opaque objects.
Resources offering this technique
Neutron Imaging
Uses hydrogen/deuterium contrast and nondestructive, high-penetrating neutrons to study a wide range of hierarchical and complex biological materials, including plant and fungal interactions, soil pore structure, and fluid transport.
Resources offering this technique
Neutron Macromolecular Crystallography
Provides information about the location of critical hydrogen atoms in protein crystals at atomic resolution. This enables studies of hydrogen bonding networks and protonation states of catalytic residues.
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Neutron Spectroscopy
Provides information about biomolecular atomic motions in time and space.
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Small-Angle Neutron Scattering
Uses differential neutron scattering of hydrogen and deuterium to study complex ensemble structures of biological materials.
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Soft X-Ray Tomography
A non-invasive, three-dimensional imaging technique that can measure volumes, surfaces, interfaces, membranes, and organelle connectivity within intact cells.
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Solution X-Ray Scattering (SAXS)
Characterizes macromolecular structure and behavior in solution, serving as an ideal assessment tool in iterative macromolecular engineering.
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Synchrotron Infrared Hyperspectral Imaging (sFTIR)
Correlates infrared maps with visible microscopy images to map the distributions of molecular compositions within biological samples and to reveal morphology and structure.
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X-Ray Absorption and Emission Spectroscopy
Synchrotron-based X-ray spectroscopy provides a powerful and synergistic toolkit to explore metal interactions within biological and biogeochemical systems and with the environment.
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X-Ray Fluorescence Imaging
Maps the distributions of elements and chemical species of interest within biological samples.
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X-Ray Footprinting
Identifies solvent-accessible regions of proteins and nucleic acids, indicating macromolecular binding surfaces or areas of conformational movement.
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X-Ray Macromolecular Crystallography
Uses X-rays to determine the atomic-level structures of biological molecules across a broad range of sample sizes and complexities.
Resources offering this technique
X-Ray Ptychography
Provides high-resolution imaging beyond X-ray lens limits.
Resources offering this technique
Structural Biology Techniques and Length Scales: Courtesy Stanford Synchrotron Radiation Lightsource at SLAC National Accelerator Laboratory. Individual images left to right: (1) Van Stappen, C., et al. 2019. “Spectroscopic Description of the E1 State of Mo Nitrogenase Based on Mo and Fe X-Ray Absorption and Mössbauer Studies,” Inorganic Chemistry 58(18), 12365–376. DOI:10.1021/acs.inorgchem.9b01951. Reprinted under a Creative Commons License (CC BY 4.0). (2) Kim, Y., et al. 2021. “Tipiracil Binds to Uridine Site and Inhibits Nsp15 Endoribonuclease NendoU from SARS-CoV-2,” Communications Biology 4, 193. DOI:10.1038/s42003-021-01735-9. Reprinted under a Creative Commons License (CC BY 4.0). (3) PDB ID: 6MOR. Roh, S. H., et al. 2020. “Cryo-EM and MD Infer Water-Mediated Proton Transport and Autoinhibition Mechanisms of Vo Complex,” Science Advances 6(41), eabb9605. DOI:10.1126/sciadv.abb9605. (4) Courtesy Thomas SpleIstoesser, www.scistyle.com. See also Vandavasi, V. G., et al. 2016. “A Structural Study of CESA1 Catalytic Domain of Arabidopsis Cellulose Synthesis Complex: Evidence for CESA Trimers,” Plant Physiology 170(1), 123–35. DOI:0.1104/pp.15.01356. (5) Reprinted with permission from Roth, M. S., et al. 2017. “Chromosome-Level Genome Assembly and Transcriptome of the Green Alga Chromochloris zofingiensis Illuminates Astaxanthin Production,” Proceedings of the National Academy of Sciences USA 114(21), E4296–4305. DOI:10.1073/pnas.1619928114. (6) Martin, M. C., et al. 2013. “3D Spectral Imaging with Synchrotron Fourier Transform Infrared Spectro-Microtomography,” Nature Methods 10, 861–64. DOI:10.1038/nmeth.2596. (7) Seyfferth, A. L, et al. 2017. “Evidence for the Root-Uptake of Arsenite at Lateral Root Junctions and Root Apices in Rice (Oryza sativa L.),” Soils 1(1), 3. DOI:10.3390/soils1010003. Reprinted under a Creative Commons License (CC BY 4.0). (8) Courtesy Oak Ridge National Laboratory. See also Dhiman, I., et al. 2017. “Quantifying Root Water Extraction After Drought Recovery Using sub-mm In Situ Empirical Data,” Plant Soil 417, 1–17. DOI:10.1007/s11104-017-3408-5.