[{"@context":"http:\/\/schema.org\/","@type":"BlogPosting","@id":"https:\/\/wiki.edu.vn\/en\/wiki41\/cryogenic-electron-microscopy-wikipedia\/#BlogPosting","mainEntityOfPage":"https:\/\/wiki.edu.vn\/en\/wiki41\/cryogenic-electron-microscopy-wikipedia\/","headline":"Cryogenic electron microscopy – Wikipedia","name":"Cryogenic electron microscopy – Wikipedia","description":"before-content-x4 Form of transmission electron microscopy (TEM) Titan Krios University of Leeds after-content-x4 Cryogenic electron microscopy (cryo-EM) is a cryomicroscopy","datePublished":"2016-03-19","dateModified":"2016-03-19","author":{"@type":"Person","@id":"https:\/\/wiki.edu.vn\/en\/wiki41\/author\/lordneo\/#Person","name":"lordneo","url":"https:\/\/wiki.edu.vn\/en\/wiki41\/author\/lordneo\/","image":{"@type":"ImageObject","@id":"https:\/\/secure.gravatar.com\/avatar\/c9645c498c9701c88b89b8537773dd7c?s=96&d=mm&r=g","url":"https:\/\/secure.gravatar.com\/avatar\/c9645c498c9701c88b89b8537773dd7c?s=96&d=mm&r=g","height":96,"width":96}},"publisher":{"@type":"Organization","name":"Enzyklop\u00e4die","logo":{"@type":"ImageObject","@id":"https:\/\/wiki.edu.vn\/wiki4\/wp-content\/uploads\/2023\/08\/download.jpg","url":"https:\/\/wiki.edu.vn\/wiki4\/wp-content\/uploads\/2023\/08\/download.jpg","width":600,"height":60}},"image":{"@type":"ImageObject","@id":"https:\/\/upload.wikimedia.org\/wikipedia\/commons\/thumb\/f\/fc\/Titan_Krios_University_of_Leeds.jpg\/220px-Titan_Krios_University_of_Leeds.jpg","url":"https:\/\/upload.wikimedia.org\/wikipedia\/commons\/thumb\/f\/fc\/Titan_Krios_University_of_Leeds.jpg\/220px-Titan_Krios_University_of_Leeds.jpg","height":"330","width":"220"},"url":"https:\/\/wiki.edu.vn\/en\/wiki41\/cryogenic-electron-microscopy-wikipedia\/","wordCount":8303,"articleBody":"     (adsbygoogle = window.adsbygoogle || []).push({});before-content-x4Form of transmission electron microscopy (TEM)  Titan Krios University of Leeds       (adsbygoogle = window.adsbygoogle || []).push({});after-content-x4Cryogenic electron microscopy (cryo-EM) is a cryomicroscopy technique applied on samples cooled to cryogenic temperatures. For biological specimens, the structure is preserved by embedding in an environment of vitreous ice. An aqueous sample solution is applied to a grid-mesh and plunge-frozen in liquid ethane or a mixture of liquid ethane and propane.[1] While development of the technique began in the 1970s, recent advances in detector technology and software algorithms have allowed for the determination of biomolecular structures at near-atomic resolution.[2] This has attracted wide attention to the approach as an alternative to X-ray crystallography or NMR spectroscopy for macromolecular structure determination without the need for crystallization.[3]In 2017, the Nobel Prize in Chemistry was awarded to Jacques Dubochet, Joachim Frank, and Richard Henderson “for developing cryo-electron microscopy for the high-resolution structure determination of biomolecules in solution.”[4]Nature Methods also named cryo-EM as the “Method of the Year” in 2015.[5]       (adsbygoogle = window.adsbygoogle || []).push({});after-content-x4The Federal Institute of Technology, the University of Lausanne and the University of Geneva opened the Dubochet Center For Imaging (DCI) at the end of November 2021, for the purposes of applying and further developing cryo-EM.[6] Less than a month after the first identification of the SARS-CoV-2 Omicron variant, researchers at the DCI were able to define its structure, identify the crucial mutations to circumvent individual vaccines and provide insights for new therapeutic approaches.[7]  Cryogenic electron microscopy workflowCryogenic transmission electron microscopy (cryo-TEM) is a transmission electron microscopy technique that is used in structural biology and materials science.Cryogenic transmission electron microscopy (cryoTEM) image of an intact ARMAN cell from an Iron Mountain biofilm. Image width is 576\u00a0nm.       (adsbygoogle = window.adsbygoogle || []).push({});after-content-x4Cryo-electron micrograph of the CroV giant marine virus(scale bar represents 200 nm)[13]Table of ContentsHistory[edit]Early development[edit]2017 Nobel Prize in Chemistry[edit]Potential rival to X-ray crystallography[edit]Correlative light Cryo-TEM and Cryo-ET[edit]Scanning electron cryomicroscopy[edit]See also[edit]References[edit]History[edit]Early development[edit]In the 1960s, the use of transmission electron microscopy for structure determination methods was limited because of the radiation damage due to high energy electron beams.  Scientists hypothesized that examining specimens at low temperatures would reduce beam-induced radiation damage.[14] Both liquid helium (\u2212269\u00a0\u00b0C or 4\u00a0K or \u2212452.2\u00a0\u00b0F) and liquid nitrogen (\u2212195.79\u00a0\u00b0C or 77\u00a0K or \u2212320\u00a0\u00b0F) were considered as cryogens. In 1980, Erwin Knapek and Jacques Dubochet published comments on beam damage at cryogenic temperatures sharing observations that:Thin crystals mounted on carbon film were found to be from 30 to 300 times more beam-resistant at 4\u00a0K than at room temperature… Most of our results can be explained by assuming that cryoprotection in the region of 4\u00a0K is strongly dependent on the temperature.[15]However, these results were not reproducible and amendments were published in Nature just two years later informing that the beam resistance was less significant than initially anticipated. The protection gained at 4\u00a0K was closer to “tenfold for standard samples of L-valine”,[16] than what was previously stated.In 1981, Alasdair McDowall and Jacques Dubochet, scientists at the European Molecular Biology Laboratory, reported the first successful implementation of cryo-EM.[17] McDowall and Dubochet vitrified pure water in a thin film by spraying it onto a hydrophilic carbon film that was rapidly plunged into cryogen (liquid propane or liquid ethane cooled to 77\u00a0K).  The thin layer of amorphous ice was less than 1\u00a0\u00b5m thick and an electron diffraction pattern confirmed the presence of amorphous\/vitreous ice.  In 1984, Dubochet’s group demonstrated the power of cryo-EM in structural biology with analysis of vitrified adenovirus type 2, T4 bacteriophage, Semliki Forest virus, Bacteriophage CbK, and Vesicular-Stomatitis-Virus.[18]2017 Nobel Prize in Chemistry[edit]In recognition of the impact cryo-EM has had on biochemistry, three scientists, Jacques Dubochet, Joachim Frank and Richard Henderson, were awarded the Nobel Prize in Chemistry “for developing cryo-electron microscopy for the high-resolution structure determination of biomolecules in solution.”[4]Potential rival to X-ray crystallography[edit]Traditionally, X-ray crystallography has been the most popular technique for determining the 3D structures of biological molecules.[19] However, the aforementioned improvements in cryo-EM have increased its popularity as a tool for examining the details of biological molecules. As a comparison, X-ray crystallography has been used to determine the 3D structures of 169,077 biological molecules (as of September 30, 2022) while cryo-EM has been used to determine fewer biological molecules at 12,647.[20]However, according to Nature, advancements in direct electron detectors (often referred to as a direct detection devices or DDDs) at the University of Cambridge[21] and automation of sample production by SPT labtech[22] has led to an increase in use in biological fields,[23] making cryo-EM a potential rival.The resolution of X-ray crystallography is limited by crystal purity,[24] and coaxing biological molecules into a crystalline state can be very time-consuming, taking up to months or even years.[23] Although sample preparation for cryo-EM is still laborious,[25] it does not have these issues as it does not require the sample to be form a crystal, rather samples for cryo-EM are flash-frozen and examined in their near-native states.[26]According to Proteopedia, the median resolution achieved by X-ray crystallography (as of May 19, 2019) on the Protein Data Bank is 2.05 \u00c5,[24] and the highest resolution achieved on record (as of September 30, 2022) is 0.48 \u00c5.[27] As of 2020, the majority of the protein structures determined by cryo-EM are at a lower resolution of 3\u20134 \u00c5.[28]  However, as of 2020, the best cryo-EM resolution has been recorded at 1.22 \u00c5,[25] making it a competitor in resolution in some cases.Correlative light Cryo-TEM and Cryo-ET[edit]In 2019, correlative light Cryo-TEM and Cryo-ET were used to observe tunnelling nanotubes (TNTs) in neuronal cells.[29]Scanning electron cryomicroscopy[edit]Scanning electron cryomicroscopy (cryoSEM) is a scanning electron microscopy technique with a scanning electron microscope’s cold stage in a cryogenic chamber.See also[edit]References[edit]^ Tivol WF, Briegel A, Jensen GJ (October 2008). “An improved cryogen for plunge freezing”. Microscopy and Microanalysis. 14 (5): 375\u2013379. Bibcode:2008MiMic..14..375T. doi:10.1017\/S1431927608080781. PMC\u00a03058946. PMID\u00a018793481.^ Cheng Y, Grigorieff N, Penczek PA, Walz T (April 2015). “A primer to single-particle cryo-electron microscopy”. Cell. 161 (3): 438\u2013449. doi:10.1016\/j.cell.2015.03.050. PMC\u00a04409659. PMID\u00a025910204.^ Stoddart C (1 March 2022). “Structural biology: How proteins got their close-up”. Knowable Magazine. doi:10.1146\/knowable-022822-1. Retrieved 25 March 2022.^ a b “The Nobel Prize in Chemistry 2017”. NobelPrize.org. Retrieved 2022-09-30.^ Doerr A (January 2017). “Cryo-electron tomography”. Nature Methods. 14 (1): 34. doi:10.1038\/nmeth.4115. ISSN\u00a01548-7091. S2CID\u00a027162203.^ “Inauguration of the Dubochet Center for Imaging (DCI) on the campuses of UNIGE, UNIL and EPFL”. unige.ch. 2021-11-30. Retrieved 2022-04-30.^ “Scientists uncover Omicron variant mysteries using microscopes”. swissinfo.ch. 2021-12-30. Retrieved 2022-04-30.^ Nannenga BL, Shi D, Leslie AG, Gonen T (September 2014). “High-resolution structure determination by continuous-rotation data collection in MicroED”. Nature Methods. 11 (9): 927\u2013930. doi:10.1038\/nmeth.3043. PMC\u00a04149488. PMID\u00a025086503.^ Jones CG, Martynowycz MW, Hattne J, Fulton TJ, Stoltz BM, Rodriguez JA,  et\u00a0al. (November 2018). “The CryoEM Method MicroED as a Powerful Tool for Small Molecule Structure Determination”. ACS Central Science. 4 (11): 1587\u20131592. doi:10.1021\/acscentsci.8b00760. PMC\u00a06276044. PMID\u00a030555912.^ de la Cruz MJ, Hattne J, Shi D, Seidler P, Rodriguez J, Reyes FE,  et\u00a0al. (February 2017). “Atomic-resolution structures from fragmented protein crystals with the cryoEM method MicroED”. Nature Methods. 14 (4): 399\u2013402. doi:10.1038\/nmeth.4178. PMC\u00a05376236. PMID\u00a028192420.^ Gruene T, Wennmacher JT, Zaubitzer C, Holstein JJ, Heidler J, Fecteau-Lefebvre A,  et\u00a0al. (December 2018). “Rapid Structure Determination of Microcrystalline Molecular Compounds Using Electron Diffraction”. Angewandte Chemie. 57 (50): 16313\u201316317. doi:10.1002\/anie.201811318. PMC\u00a06468266. PMID\u00a030325568.^ Cheng Y (August 2018). “Single-particle cryo-EM-How did it get here and where will it go”. Science. 361 (6405): 876\u2013880. Bibcode:2018Sci…361..876C. doi:10.1126\/science.aat4346. PMC\u00a06460916. PMID\u00a030166484.^ Xiao, C., Fischer, M.G., Bolotaulo, D.M., Ulloa-Rondeau, N., Avila, G.A., and Suttle, C.A. (2017) “Cryo-EM reconstruction of the Cafeteria roenbergensis virus capsid suggests novel assembly pathway for giant viruses”. Scientific Reports, 7: 5484. doi:10.1038\/s41598-017-05824-w.^ Dubochet J, Knapek E (April 2018). “Ups and downs in early electron cryo-microscopy”. PLOS Biology. 16 (4): e2005550. doi:10.1371\/journal.pbio.2005550. PMC\u00a05929567. PMID\u00a029672565.^ Knapek E, Dubochet J (August 1980). “Beam damage to organic material is considerably reduced in cryo-electron microscopy”. Journal of Molecular Biology. 141 (2): 147\u2013161. doi:10.1016\/0022-2836(80)90382-4. PMID\u00a07441748.^ Newmark P (30 September 1982). “Cryo-transmission microscopy Fading hopes”. Nature. 299 (5882): 386\u2013387. Bibcode:1982Natur.299..386N. doi:10.1038\/299386c0.^ Dubochet J, McDowall AW (December 1981). “Vitrification of Pure Water for Electron Microscopy”. Journal of Microscopy. 124 (3): 3\u20134. doi:10.1111\/j.1365-2818.1981.tb02483.x.^ Adrian M, Dubochet J, Lepault J, McDowall AW (March 1984). “Cryo-electron microscopy of viruses” (PDF). Nature. 308 (5954): 32\u201336. Bibcode:1984Natur.308…32A. doi:10.1038\/308032a0. PMID\u00a06322001. S2CID\u00a04319199.^ Smyth MS, Martin JH (February 2000). “x ray crystallography”. Molecular Pathology. 53 (1): 8\u201314. doi:10.1136\/mp.53.1.8. PMC\u00a01186895. PMID\u00a010884915.^ “RCSB PDB Statistics”. www.rcsb.org. Retrieved 2020-09-30.^ Callaway E (September 2015). “The revolution will not be crystallized: a new method sweeps through structural biology”. Nature. 525 (7568): 172\u2013174. Bibcode:2015Natur.525..172C. doi:10.1038\/525172a. PMID\u00a026354465.^ Baker M (September 2018). “Cryo-electron microscopy shapes up”. Nature. 561 (7724): 565\u2013567. Bibcode:2018Natur.561..565B. doi:10.1038\/d41586-018-06791-6. PMID\u00a030254359.^ a b Callaway E (February 2020). “Revolutionary cryo-EM is taking over structural biology”. Nature. 578 (7794): 201. Bibcode:2020Natur.578..201C. doi:10.1038\/d41586-020-00341-9. PMID\u00a032047310.^ a b “Resolution – Proteopedia, life in 3D”. proteopedia.org. Retrieved 2020-10-27.^ a b Nakane T, Kotecha A, Sente A, McMullan G, Masiulis S, Brown PM,  et\u00a0al. (November 2020). “Single-particle cryo-EM at atomic resolution”. Nature. 587 (7832): 152\u2013156. Bibcode:2020Natur.587..152N. doi:10.1038\/s41586-020-2829-0. PMC\u00a07611073. PMID\u00a033087931.^ Wang HW, Wang JW (2016-08-20). “How cryo-electron microscopy and X-ray crystallography complement each other”. Protein Science. 26 (1): 32\u201339. doi:10.1002\/pro.3022. PMC\u00a05192981. PMID\u00a027543495.^ Schmidt A, Teeter M, Weckert E, Lamzin VS (April 2011). “Crystal structure of small protein crambin at 0.48 \u00c5 resolution”. Acta Crystallographica. Section F, Structural Biology and Crystallization Communications. 67 (Pt 4): 424\u2013428. doi:10.1107\/S1744309110052607. PMC\u00a03080141. PMID\u00a021505232.^ Yip KM, Fischer N, Paknia E, Chari A, Stark H (November 2020). “Atomic-resolution protein structure determination by cryo-EM”. Nature. 587 (7832): 157\u2013161. Bibcode:2020Natur.587..157Y. doi:10.1038\/s41586-020-2833-4. PMID\u00a033087927. S2CID\u00a0224823207.^ Sartori-Rupp A, Cordero Cervantes D, Pepe A, Gousset K, Delage E, Corroyer-Dulmont S,  et\u00a0al. (January 2019). “Correlative cryo-electron microscopy reveals the structure of TNTs in neuronal cells”. Nature Communications. 10 (1): 342. Bibcode:2019NatCo..10..342S. doi:10.1038\/s41467-018-08178-7. PMC\u00a06341166. PMID\u00a030664666.     (adsbygoogle = window.adsbygoogle || []).push({});after-content-x4"},{"@context":"http:\/\/schema.org\/","@type":"BreadcrumbList","itemListElement":[{"@type":"ListItem","position":1,"item":{"@id":"https:\/\/wiki.edu.vn\/en\/wiki41\/#breadcrumbitem","name":"Enzyklop\u00e4die"}},{"@type":"ListItem","position":2,"item":{"@id":"https:\/\/wiki.edu.vn\/en\/wiki41\/cryogenic-electron-microscopy-wikipedia\/#breadcrumbitem","name":"Cryogenic electron microscopy – Wikipedia"}}]}]