The hottest naturemethods annual technical single

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Naturemethods annual technology: near the end of the year, major magazines carried out a year-end inventory successively. On December 30, naturemethods also conducted an annual technology inventory and selected the most popular and widely influential technology in 2015. 2. Standard configuration result: cryo em

the three-dimensional structure of a protein or protein complex can provide important insights into its biological functions. As a structure determination technology, single particle cryo EM ranks only behind the new product of high-resolution methods such as X-ray crystallography and nuclear magnetic resonance (NMR) spectroscopy. Due to the technological progress in recent years, cryo em can now be used to analyze the near atomic resolution structure, which is rapidly changing

for decades, X-ray crystallography has been the preferred method to analyze protein structure. However, many proteins, especially membrane proteins and protein complexes, are difficult to crystallize. Some alternative methods to traditional crystallography have their own limitations. For example, serialfemtosecondcrystallography technology uses X-ray free electron laser (xfel), which no longer requires a single large protein crystal, but obtains a large number of microcrystals that are easy to generate. However, under the highly specialized xfel, the competition for beamtime is very fierce. NMR spectroscopy can be used to analyze the structure of small proteins, but it is still difficult to apply it to larger proteins

different from crystallography, cryo EM is particularly suitable for obtaining structural information of large protein complexes and some systems that display a variety of conformations or composition states. Over the past few decades, researchers in this initially small field have been making steady progress to improve the resolution of cryo EM and further expand its biological adaptability. In this issue of naturemethods, evanogales introduced the development history of cryo em as a mainstream structural biology technology

this once small field is now advancing by leaps and bounds. A new type of highly sensitive direct detection camera can directly capture electrons, which may achieve a leap in resolution. The first papers exploring these new detectors were published in 2013. In 2014, I saw several papers using cryo em to analyze some important high-resolution structures. In 2015, many studies have broken through the 3 angstrom resolution barrier - an unprecedented feat that even some long-term cryo EM practitioners were surprised

but a good detector is not everything. A successful cryo EM research largely depends on good sample preparation and complex image processing software. Allisondoerr discusses this issue in this issue of naturemethods

cryo EM resolution reform has just begun, and robertglaeser discussed this in a review article. The increased sensitivity of detector technology brings opportunities to develop some new and improved methods, which will further promote the improvement of resolution, applicability and ease of use. Although cryo EM is especially suitable for large protein complexes, these protein complexes studied so far are mainly easy to pick fruits. At present, there is an urgent need for some practical, repeatable and general sample preparation methods to expand the applicability of cryo EM and detect all structures that cannot be determined by structural technology so far. Data analysis methods also need further improvement. Researchers hope to get some simple and reliable calculation methods to transform the original two-dimensional image into three-dimensional protein structure, especially for detecting systems with structural heterogeneity

is the key to control and guide its biological reaction and avoid allogeneic reaction

like all scientific fields in the rapid growth period, cryo EM also has the trouble of growth. A special article in this issue of naturemethods magazine discusses this problem. It is encouraging that many countries are building national user facilities with high-end instruments. However, at present, the supply of this high-end instrument is in short supply. Many researchers hope to use this technology, but cryo EM is not an automation technology (at least not at present). There are many complex steps in the process of sample preparation and data analysis. Researchers must carefully apply, record and verify them correctly to avoid mistakes. It is essential to ensure that new cryo EM practitioners receive appropriate training. He explained: "the example of three stars is very serious

however, these developments of cryo EM do not mean the end of crystallography. X-ray crystallography will still be a powerful technology for analyzing the structure of proteins that are easy to crystallize

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