{ "id": "ep5synb011", "slug": "cryomicroscopy", "title": "Cryomicroscopy", "updated": "2026-09-30", "topic_path": [ { "id": "physical", "label": "Physical world and mathematics", "api_url": "https://www.edgepedia.org/api/v1/topics/physical" }, { "id": "physical.physics", "label": "Physics", "api_url": "https://www.edgepedia.org/api/v1/topics/physical.physics" }, { "id": "physical.physics.practice", "label": "Physics methods, practice, and community", "api_url": "https://www.edgepedia.org/api/v1/topics/physical.physics.practice" }, { "id": "physical.physics.practice.optical-and-light-microscopy", "label": "Optical and light microscopy", "api_url": "https://www.edgepedia.org/api/v1/topics/physical.physics.practice.optical-and-light-microscopy" } ], "geo": [ { "id": "geo.nongeo.t1946.physical.physics.practice", "label": "Non-geographic · 1946 to 2000: Physics methods, practice, and community", "api_url": "https://www.edgepedia.org/api/v1/geo/geo.nongeo.t1946.physical.physics.practice", "path": [ { "id": "geo.nongeo", "label": "Non-geographic", "api_url": "https://www.edgepedia.org/api/v1/geo/geo.nongeo" }, { "id": "geo.nongeo.t1946", "label": "Non-geographic · 1946 to 2000", "api_url": "https://www.edgepedia.org/api/v1/geo/geo.nongeo.t1946" }, { "id": "geo.nongeo.t1946.physical", "label": "Physical world and mathematics", "api_url": "https://www.edgepedia.org/api/v1/geo/geo.nongeo.t1946.physical" }, { "id": "geo.nongeo.t1946.physical.physics", "label": "Physics", "api_url": "https://www.edgepedia.org/api/v1/geo/geo.nongeo.t1946.physical.physics" }, { "id": "geo.nongeo.t1946.physical.physics.practice", "label": "Physics methods, practice, and community", "api_url": "https://www.edgepedia.org/api/v1/geo/geo.nongeo.t1946.physical.physics.practice" } ] }, { "id": "geo.nongeo.t2001.physical.physics.practice", "label": "Non-geographic · 2001 to 2020: Physics methods, practice, and community", "api_url": "https://www.edgepedia.org/api/v1/geo/geo.nongeo.t2001.physical.physics.practice", "path": [ { "id": "geo.nongeo", "label": "Non-geographic", "api_url": "https://www.edgepedia.org/api/v1/geo/geo.nongeo" }, { "id": "geo.nongeo.t2001", "label": "Non-geographic · 2001 to 2020", "api_url": "https://www.edgepedia.org/api/v1/geo/geo.nongeo.t2001" }, { "id": "geo.nongeo.t2001.physical", "label": "Physical world and mathematics", "api_url": "https://www.edgepedia.org/api/v1/geo/geo.nongeo.t2001.physical" }, { "id": "geo.nongeo.t2001.physical.physics", "label": "Physics", "api_url": "https://www.edgepedia.org/api/v1/geo/geo.nongeo.t2001.physical.physics" }, { "id": "geo.nongeo.t2001.physical.physics.practice", "label": "Physics methods, practice, and community", "api_url": "https://www.edgepedia.org/api/v1/geo/geo.nongeo.t2001.physical.physics.practice" } ] } ], "excerpt": "Cryomicroscopy images samples held at cryogenic temperatures, usually thin films of vitreous ice, to preserve native structure. Its three main modalities are single-particle analysis, electron tomography, and microelectron diffraction.", "snippet": "Cryomicroscopy images samples held at cryogenic temperatures, usually thin films of vitreous ice, to preserve native structure. Its three main modalities are single-particle analysis, electron tomography, and microelectron diffraction.", "node": "physical.physics.practice.optical-and-light-microscopy", "markdown": "# Cryomicroscopy\n\nCryomicroscopy images samples held at cryogenic temperatures, usually as thin films of vitreous ice, to preserve native or frozen structure for imaging. Its three main modalities are single-particle analysis of purified macromolecules, electron tomography of cells in their native state, and microelectron diffraction of microcrystals.[1](https://link.springer.com/article/10.1186/s44398-025-00017-2) The best single-particle resolution, mouse apoferritin, reaches 1.09 Å.[2](https://pmc.ncbi.nlm.nih.gov/articles/PMC12403223/) Cryo-electron microscopy is used for the high-resolution structure determination of biomolecules in solution.[3](https://www.nobelprize.org/uploads/2018/06/advanced-chemistryprize2017-1.pdf)\n\n| Fact | Value |\n|---|---|\n| Main modalities | Single-particle analysis, electron tomography, microelectron diffraction [1](https://link.springer.com/article/10.1186/s44398-025-00017-2) |\n| Vitrification on plunging | Aqueous film cools to ~90 K in under 0.1 ms; maximum vitrifiable layer ~1 µm [4](https://www.cambridge.org/core/journals/quarterly-reviews-of-biophysics/article/current-limitations-to-highresolution-structure-determination-by-singleparticle-cryoem/43D0BBE24C3D620D0CC999C61FDCB48E) |\n| Devitrification on warming | Reported at about −140 °C [3](https://www.nobelprize.org/uploads/2018/06/advanced-chemistryprize2017-1.pdf) and around −145 °C [5](https://www.sciencedirect.com/science/article/pii/S0968432824001628) |\n| Cryoprotection | Beam damage reduced 30–300-fold versus room temperature [6](https://journals.plos.org/plosbiology/article?id=10.1371%2Fjournal.pbio.2005550) |\n| Best resolutions | 1.09 Å (single-particle apoferritin) [2](https://pmc.ncbi.nlm.nih.gov/articles/PMC12403223/); 2–4 nm routine for cellular cryo-ET [7](https://www.osti.gov/servlets/purl/3020406) |\n| Typical dose | 30–40 e⁻/Ų per dataset [4](https://www.cambridge.org/core/journals/quarterly-reviews-of-biophysics/article/current-limitations-to-highresolution-structure-determination-by-singleparticle-cryoem/43D0BBE24C3D620D0CC999C61FDCB48E), corresponding to roughly 90–120 MGy at 3 MGy per 1 e⁻/Ų [8](https://www.sciencedirect.com/science/article/pii/S0959440X25000235) |\n| Sample thickness | Under 300 Å ice for electron transparency [2](https://pmc.ncbi.nlm.nih.gov/articles/PMC12403223/); cellular lamellae milled to ~0.2 µm [7](https://www.osti.gov/servlets/purl/3020406) |\n\n## How it works\n\n**Vitrification is the physical core.** When a thin aqueous film on an electron-microscopy grid plunges into liquid ethane, it cools from room temperature to about 90 K in less than 0.1 ms, faster than ice crystals can nucleate, so the water solidifies as amorphous vitreous ice.[4](https://www.cambridge.org/core/journals/quarterly-reviews-of-biophysics/article/current-limitations-to-highresolution-structure-determination-by-singleparticle-cryoem/43D0BBE24C3D620D0CC999C61FDCB48E) The freezing rates required, on the order of 10,000 °C per second, are achievable at atmospheric pressure only for layers thinner than about 20 µm.[9](https://www.colorado.edu/facility/ems/sites/default/files/attached-files/mmanual.pdf) On warming, amorphous ice crystallizes: the Nobel background gives about −140 °C[3](https://www.nobelprize.org/uploads/2018/06/advanced-chemistryprize2017-1.pdf) and a methods study around −145 °C,[5](https://www.sciencedirect.com/science/article/pii/S0968432824001628) so specimens must stay well below these temperatures throughout transfer and imaging.\n\nCooling also protects against the electron beam itself, which destroys biological macromolecules at the same time it produces the image.[10](https://www.annualreviews.org/content/journals/10.1146/annurev-biophys-070317-032828) Knapek and Dubochet measured a 30–300-fold reduction in damage relative to room temperature.[6](https://journals.plos.org/plosbiology/article?id=10.1371%2Fjournal.pbio.2005550) An earlier report that damage is \"dramatically reduced\" near 4.2 K in a superconducting-lens microscope[11](https://doi.org/10.1038/277380a0) was later reevaluated: no certain further cryoprotection is observed near 4 K compared with liquid-nitrogen temperature.[6](https://journals.plos.org/plosbiology/article?id=10.1371%2Fjournal.pbio.2005550) Contrast sets a thickness limit: the mean free path of 300 kV electrons in water is roughly 300 nm, and ice thinner than about 300 Å gives the best transparency.[4](https://www.cambridge.org/core/journals/quarterly-reviews-of-biophysics/article/current-limitations-to-highresolution-structure-determination-by-singleparticle-cryoem/43D0BBE24C3D620D0CC999C61FDCB48E) • [2](https://pmc.ncbi.nlm.nih.gov/articles/PMC12403223/)\n\n## How it is done\n\nA typical single-particle workflow starts with a 3–4 µL droplet of sample on an EM grid, blotted with filter paper to leave a thin liquid film, then plunged into cryogen.[5](https://www.sciencedirect.com/science/article/pii/S0968432824001628) Blotting is irreproducible because filter paper contact varies, so blot-free microfluidic systems (SPT [Chameleon](https://www.edgepedia.org/chameleon), CryoSol Vitrojet, cryoWriter) vitrify within milliseconds and shorten exposure to the air–water interface.[2](https://pmc.ncbi.nlm.nih.gov/articles/PMC12403223/) Liquid ethane must be held below the amorphous-to-crystalline transition near −145 °C, and blot time strongly affects ice quality: in one test at a 45 °C chamber, 1.5–2.5 s gave consistent amorphous ice, under 1 s gave thick opaque ice, and 4 s left grids empty.[5](https://www.sciencedirect.com/science/article/pii/S0968432824001628)\n\nFor thick samples, high-pressure freezing applies about 2,050 bar immediately before freezing; one methods manual puts well-frozen thickness at up to 0.6 mm,[9](https://www.colorado.edu/facility/ems/sites/default/files/attached-files/mmanual.pdf) while a cryo-ET survey gives 10–200 µm.[7](https://www.osti.gov/servlets/purl/3020406) Because TEM samples must be thinner than about 0.3–0.5 µm, cellular samples are milled by cryo-focused ion beam into ~0.2 µm lamellae, which extends access to cells and tissues.[7](https://www.osti.gov/servlets/purl/3020406) Support films matter: all-gold grids reduce beam-induced specimen movement to below 1 Å,[12](https://doi.org/10.1126/science.abb7927) and graphene-oxide films provide a robust preparation surface.[13](https://doi.org/10.1016/j.jsb.2018.07.007) On a 300 kV Titan Krios with a K3 detector, faster optical centering collects 736–758 movies per hour, and final datasets run 2,000–8,000 movies.[14](https://link.springer.com/article/10.1186/s44330-025-00025-8) Processing uses packages such as RELION[15](https://doi.org/10.1016/j.jsb.2012.09.006) and cryoSPARC;[16](https://doi.org/10.1038/nmeth.4169) published protocols carry the practitioner from purified complex in aqueous solution to micrographs suitable for 3D structure determination.[17](https://europepmc.org/article/pmc/5140023)\n\n## Origin\n\nEarlier work used windowless, differentially pumped hydration stages to keep samples hydrated at room temperature in the microscope.[18](https://pmc.ncbi.nlm.nih.gov/articles/PMC3291472/) [Kenneth A. Taylor](https://www.edgepedia.org/kenneth-a-taylor) and Robert M. Glaeser reported electron diffraction of frozen, hydrated protein crystals in *Science* in 1974,[19](https://doi.org/10.1126/science.186.4168.1036) showing diffraction beyond 3 Å from frozen catalase crystals and that hydration is maintained at cryogenic temperatures.[3](https://www.nobelprize.org/uploads/2018/06/advanced-chemistryprize2017-1.pdf) Peter Brüggeller and [Erwin Mayer](https://www.edgepedia.org/erwin-mayer) reported complete vitrification of pure liquid water in 1980.[20](https://doi.org/10.1038/288569a0) In December 1981 the *Journal of Microscopy* published Dubochet and McDowall's short paper on vitrification of pure water for electron microscopy,[21](https://doi.org/10.1111/j.1365-2818.1981.tb02483.x) • [22](https://onlinelibrary.wiley.com/doi/10.1111/j.1365-2818.2011.03569.x) in which water sprayed on a carbon film was rapidly immersed in liquid ethane or propane at about −190 °C.[3](https://www.nobelprize.org/uploads/2018/06/advanced-chemistryprize2017-1.pdf) Adrian, Dubochet, Lepault, and McDowall's \"Cryo-electron microscopy of viruses\" (*Nature*, 1984) prepared thin unsupported vitrified water layers and is described as the article with the most impact, paving the way for icosahedral virus structures.[23](https://doi.org/10.1038/308032a0) • [18](https://pmc.ncbi.nlm.nih.gov/articles/PMC3291472/) Dubochet and colleagues' 1988 review in *Quarterly Reviews of Biophysics* consolidated the method,[24](https://doi.org/10.1017/s0033583500004297) and an EMBO course spread the technique rapidly.[25](https://www.cambridge.org/core/journals/quarterly-reviews-of-biophysics/article/abs/cryoelectron-microscopy-of-vitrified-specimens/471BB7106E820E9B88F1FAED9875C277) A single-particle reconstruction reconstructed the Semliki Forest virus envelope from cryo-micrographs.[26](https://doi.org/10.1038/320533a0) High-resolution structures could be obtained by averaging many copies, from bacteriorhodopsin 2D crystals.[3](https://www.nobelprize.org/uploads/2018/06/advanced-chemistryprize2017-1.pdf) Noncrystalline samples first reached near 3 Å in 2014, the \"resolution revolution\"; cryo-EM was [Nature Methods](https://www.edgepedia.org/nature-methods)' Method of the Year 2015, and the 2017 Nobel Prize followed.[1](https://link.springer.com/article/10.1186/s44398-025-00017-2)\n\n## Variants\n\n**Single-particle cryo-EM** averages thousands of identical particles; hemoglobin (64 kDa) has been determined at 3.2 Å from about 175,000 particles.[10](https://www.annualreviews.org/content/journals/10.1146/annurev-biophys-070317-032828) A Volta phase plate provides enough contrast to identify 64-kDa particles at defocus below 500 nm.[10](https://www.annualreviews.org/content/journals/10.1146/annurev-biophys-070317-032828) **Cryo-electron tomography** images the cellular interior in near-native state; cryogenic focused ion beam fabrication of thin lamellae is the enabling step,[27](https://www.nature.com/articles/s41592-023-01783-5) and cryo-FIB lift-out brings molecular-resolution tomography to native *Caenorhabditis elegans* tissue.[28](https://doi.org/10.1038/s41592-019-0497-5) [Tomography](https://www.edgepedia.org/tomography) requires sample thickness below about 300 nm.[29](https://pubs.aip.org/aca/sdy/article/12/2/020901/3344539/Unraveling-atomic-complexity-from-frozen-samples) **MicroED** collects electron diffraction from micro- and nanocrystals a billionth the size needed for conventional [X-ray diffraction](https://www.edgepedia.org/x-ray-diffraction).[30](https://doi.org/10.1038/s41592-019-0395-x) • [29](https://pubs.aip.org/aca/sdy/article/12/2/020901/3344539/Unraveling-atomic-complexity-from-frozen-samples) **Cryo-fluorescence microscopy** locates fluorescent markers in vitrified samples before EM study, and photobleaching is slower at −140 °C.[31](https://onlinelibrary.wiley.com/doi/10.1111/j.1365-2818.2007.01794.x)\n\nOn hardware, new 100 kV systems minimize radiation damage while reporting resolution comparable to 200 and 300 kV instruments,[2](https://pmc.ncbi.nlm.nih.gov/articles/PMC12403223/) though their advantage fades with thick ice, and 50–200 nm lamellae are too thick for them.[32](https://doi.org/10.1016/j.str.2025.07.007) Machine-learning tools now cover the pipeline: CryoDRGN for heterogeneous reconstruction,[33](https://doi.org/10.1038/s41592-020-01049-4) ModelAngelo for automated model building,[34](https://doi.org/10.1038/s41586-024-07215-4) and AreTomo[35](https://doi.org/10.1016/j.yjsbx.2022.100068) with nextPYP[36](https://doi.org/10.1038/s41592-023-02045-0) for tomographic alignment.\n\n## Applications\n\nSingle-particle cryo-EM is used across structural biology for complexes from 50 kDa to 5 MDa,[4](https://www.cambridge.org/core/journals/quarterly-reviews-of-biophysics/article/current-limitations-to-highresolution-structure-determination-by-singleparticle-cryoem/43D0BBE24C3D620D0CC999C61FDCB48E) including structure-based drug discovery.[2](https://pmc.ncbi.nlm.nih.gov/articles/PMC12403223/) Cellular cryo-ET on FIB lamellae supports in situ structural cell biology, with more in situ structures obtained at increasingly higher resolution as microscopes, detectors, and processing improve.[27](https://www.nature.com/articles/s41592-023-01783-5) As of May 2024, more than 35,000 structure entries had been deposited in the Electron Microscopy Data Bank.[29](https://pubs.aip.org/aca/sdy/article/12/2/020901/3344539/Unraveling-atomic-complexity-from-frozen-samples) Structure-prediction methods such as [AlphaFold](https://www.edgepedia.org/alphafold) serve as a complementary source of models for interpretation.[37](https://doi.org/10.1038/s41586-021-03819-2)\n\n## Limitations and alternatives\n\n**Radiation damage is unavoidable.** At 300 keV the overall B-factor grows by about 5 Ų per 1 e⁻/Ų of fluence, and most specimens receive about 60 MGy; after the first movie frame at 1 e⁻/Ų (3 MGy) many aspartate and glutamate side chains decarboxylate.[8](https://www.sciencedirect.com/science/article/pii/S0959440X25000235) **The air–water interface** is the other main limit: complexes collide with it thousands of times per second during preparation, and only a small fraction of particles often contribute to high-resolution maps, for example 19% for a GABA-A receptor at 3.8 Å, 15% for [P-glycoprotein](https://www.edgepedia.org/p-glycoprotein) at 3.4 Å, and 5.7% for a 4 Å sodium channel.[4](https://www.cambridge.org/core/journals/quarterly-reviews-of-biophysics/article/current-limitations-to-highresolution-structure-determination-by-singleparticle-cryoem/43D0BBE24C3D620D0CC999C61FDCB48E) Beam-induced movement, the second limiting effect, has been largely resolved by all-gold support grids.[4](https://www.cambridge.org/core/journals/quarterly-reviews-of-biophysics/article/current-limitations-to-highresolution-structure-determination-by-singleparticle-cryoem/43D0BBE24C3D620D0CC999C61FDCB48E) • [12](https://doi.org/10.1126/science.abb7927)\n\nSize limits remain: the calculated physical limit for single-particle imaging is 20–40 kDa, not yet achieved, while the average human cell protein is 35 kDa;[29](https://pubs.aip.org/aca/sdy/article/12/2/020901/3344539/Unraveling-atomic-complexity-from-frozen-samples) attempts on roughly 32 kDa targets had failed at the time of one review.[10](https://www.annualreviews.org/content/journals/10.1146/annurev-biophys-070317-032828) Cryo-tomography is hard to push beyond 3–4 nm because 50–100 low-dose images of the same area must be collected;[38](https://pubs.acs.org/doi/full/10.1021/cr100353t) a round costs roughly US$3,000–5,000 and takes 1–2 weeks, and only about 0.5–4% of cellular material is captured within lamellae.[7](https://www.osti.gov/servlets/purl/3020406) As resolution yardsticks, resolving α-helices requires better than about 9 Å and β-strands better than 4.5 Å.[38](https://pubs.acs.org/doi/full/10.1021/cr100353t)\n\nCompared with other structural methods, single-particle cryo-EM has matured over the past decade into a robust method complementing [X-ray crystallography](https://www.edgepedia.org/x-ray-crystallography) and NMR, with exponential growth in structures solved annually.[39](https://www.annualreviews.org/content/journals/10.1146/annurev-biophys-111622-091300) Dose budgets differ sharply: cryo-EM data are usually collected at 30–40 e⁻/Ų, whereas a typical X-ray crystallography dataset uses 1.5 MGy (0.5 e⁻/Ų equivalent).[4](https://www.cambridge.org/core/journals/quarterly-reviews-of-biophysics/article/current-limitations-to-highresolution-structure-determination-by-singleparticle-cryoem/43D0BBE24C3D620D0CC999C61FDCB48E) Cryo-EM starts from a purified complex in aqueous solution,[17](https://europepmc.org/article/pmc/5140023) and MicroED works with crystals a billionth the size required for conventional X-ray diffraction.[29](https://pubs.aip.org/aca/sdy/article/12/2/020901/3344539/Unraveling-atomic-complexity-from-frozen-samples)\n\n## References\n\n1. [Artificial intelligence in cryo-EM: emerging deep neural network methods from sample preparation, particle picking, map reconstruction, modelling to enhanced resolution | BMC Artificial Intelligence](https://link.springer.com/article/10.1186/s44398-025-00017-2)\n2. [Advances in cryo-electron microscopy (cryoEM) for structure-based drug discovery (PMC)](https://pmc.ncbi.nlm.nih.gov/articles/PMC12403223/)\n3. [The Development of Cryo-Electron Microscopy (Nobel Prize for Chemistry 2017, Advanced Information)](https://www.nobelprize.org/uploads/2018/06/advanced-chemistryprize2017-1.pdf)\n4. [Current limitations to high-resolution structure determination by single-particle cryoEM (Quarterly Reviews of Biophysics)](https://www.cambridge.org/core/journals/quarterly-reviews-of-biophysics/article/current-limitations-to-highresolution-structure-determination-by-singleparticle-cryoem/43D0BBE24C3D620D0CC999C61FDCB48E)\n5. [Practical considerations for plunge freezing samples over 40 °C for Cryo-EM (2024)](https://www.sciencedirect.com/science/article/pii/S0968432824001628)\n6. [Ups and downs in early electron cryo-microscopy (PLOS Biology)](https://journals.plos.org/plosbiology/article?id=10.1371%2Fjournal.pbio.2005550)\n7. [Cryogenic electron tomography by the numbers: Charting underexplored lineages in structural cell biology](https://www.osti.gov/servlets/purl/3020406)\n8. [Extending the reach of single-particle cryoEM (Current Opinion in Structural Biology, 2025)](https://www.sciencedirect.com/science/article/pii/S0959440X25000235)\n9. [High pressure freezing and freeze-substitution methods manual (University of Colorado EM Services)](https://www.colorado.edu/facility/ems/sites/default/files/attached-files/mmanual.pdf)\n10. [How Good Can Single-Particle Cryo-EM Become? What Remains Before It Approaches Its Physical Limits? (Annual Review of Biophysics)](https://www.annualreviews.org/content/journals/10.1146/annurev-biophys-070317-032828)\n11. [I. DIETRICH and colleagues (1979). Reduction of radiation damage in an electron microscope with a superconducting lens system. Nature.](https://doi.org/10.1038/277380a0)\n12. [Katerina Naydenova, Peipei Jia, Christopher J. Russo (2020). Cryo-EM with sub–1 Å specimen movement. Science.](https://doi.org/10.1126/science.abb7927)\n13. [Eugene Palovcak and colleagues (2018). A simple and robust procedure for preparing graphene-oxide cryo-EM grids. Journal of Structural Biology.](https://doi.org/10.1016/j.jsb.2018.07.007)\n14. [Efficient strategies and troubleshooting for single particle cryoEM data collection using EPU (BMC Methods)](https://link.springer.com/article/10.1186/s44330-025-00025-8)\n15. [Sjors H.W. Scheres (2012). RELION: Implementation of a Bayesian approach to cryo-EM structure determination. Journal of Structural Biology.](https://doi.org/10.1016/j.jsb.2012.09.006)\n16. [Ali Punjani and colleagues (2017). cryoSPARC: algorithms for rapid unsupervised cryo-EM structure determination. Nature Methods.](https://doi.org/10.1038/nmeth.4169)\n17. [Specimen Preparation for High-Resolution Cryo-EM (Passmore & Russo, Methods Enzymol. 579:51-86, 2016)](https://europepmc.org/article/pmc/5140023)\n18. [Retrospective on the Early Development of Cryoelectron Microscopy of Macromolecules and a Prospective on Opportunities for the Future (Glaeser & Taylor)](https://pmc.ncbi.nlm.nih.gov/articles/PMC3291472/)\n19. [Kenneth A. Taylor, Robert M. Glaeser (1974). Electron Diffraction of Frozen, Hydrated Protein Crystals. Science.](https://doi.org/10.1126/science.186.4168.1036)\n20. [Peter Brüggeller, Erwin Mayer (1980). Complete vitrification in pure liquid water and dilute aqueous solutions. Nature.](https://doi.org/10.1038/288569a0)\n21. [J. Dubochet, A.W. McDowall (1981). 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Nature.](https://doi.org/10.1038/320533a0)\n27. [Cryo-electron tomography on focused ion beam lamellae transforms structural cell biology](https://www.nature.com/articles/s41592-023-01783-5)\n28. [Miroslava Schaffer and colleagues (2019). A cryo-FIB lift-out technique enables molecular-resolution cryo-ET within native Caenorhabditis elegans tissue. Nature Methods.](https://doi.org/10.1038/s41592-019-0497-5)\n29. [Unraveling atomic complexity from frozen samples (Structural Dynamics, 2025)](https://pubs.aip.org/aca/sdy/article/12/2/020901/3344539/Unraveling-atomic-complexity-from-frozen-samples)\n30. [Brent L. Nannenga, Tamir Gonen (2019). The cryo-EM method microcrystal electron diffraction (MicroED). Nature Methods.](https://doi.org/10.1038/s41592-019-0395-x)\n31. [Cryo-fluorescence microscopy facilitates correlations between light and cryo-electron microscopy and reduces the rate of photobleaching](https://onlinelibrary.wiley.com/doi/10.1111/j.1365-2818.2007.01794.x)\n32. 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[Prospects and Limitations of High-Resolution Single-Particle Cryo-Electron Microscopy (Annual Review of Biophysics)](https://www.annualreviews.org/content/journals/10.1146/annurev-biophys-111622-091300)\n\n---\n*Topic: Encyclopedia › Physical world and mathematics › Physics › Physics methods, practice, and community › Optical and light microscopy*\n\n*Initially written Sep 29, 2026 · Reviewed: Sep 30, 2026 · Edited: Sep 30, 2026 · Last review: Sep 30, 2026*\n\n*Copyright 2026 EdgeChat AI, a subsidiary of Biostate AI.*\n\nLicense: Edgepedia Community License 1.0, https://www.edgepedia.org/edgepedia/license\n", "same_as": [], "url": "https://www.edgepedia.org/cryomicroscopy", "markdown_url": "https://www.edgepedia.org/cryomicroscopy.md", "license": { "name": "Edgepedia Community License 1.0", "url": "https://www.edgepedia.org/edgepedia/license", "summary": "Free with credit, commercial use included. AI training is open to everyone. For other uses, organizations over USD 100M in revenue or 100M monthly users license separately.", "spdx": "LicenseRef-Edgepedia-Community-1.0" }, "credit": "\"Cryomicroscopy\", Edgepedia (EdgeChat), https://www.edgepedia.org/cryomicroscopy. Edgepedia Community License 1.0.", "credit_md": "\"[Cryomicroscopy](https://www.edgepedia.org/cryomicroscopy)\", Edgepedia (EdgeChat), [https://www.edgepedia.org/cryomicroscopy](https://www.edgepedia.org/cryomicroscopy). [Edgepedia Community License 1.0](https://www.edgepedia.org/edgepedia/license).", "credit_html": "\"Cryomicroscopy\", Edgepedia (EdgeChat), https://www.edgepedia.org/cryomicroscopy. Edgepedia Community License 1.0.", "speakable": "Cryomicroscopy images samples held at cryogenic temperatures, usually thin films of vitreous ice, to preserve native structure. Its three main modalities are single-particle analysis, electron tomography, and microelectron diffraction." }