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France After gradually moving away from preparation methods prone to artefacts such as plastic embedding and negative staining for cell sections and single particles, the field of cryo electron microscopy (cryo-EM) is now heading off at unprecedented speed towards high-resolution analysis of biological objects of various sizes. This 'revolution in resolution' is happening largely thanks to new developments of new-generation cameras used for recording the images in the cryo electron microscope which have much increased sensitivity being based on complementary metal oxide semiconductor devices. Combined with advanced image processing and 3D reconstruction, the cryo-EM analysis of nucleoprotein complexes can provide unprecedented insights at molecular and atomic levels and address regulatory mechanisms in the cell. These advances reinforce the integrative role of cryo-EM in synergy with other methods such as X-ray crystallography, fluorescence imaging or focussed-ion beam milling as exemplified here by some recent studies from our laboratory on ribosomes, viruses, chromatin and nuclear receptors. Such multi-scale and multi-resolution approaches allow integrating molecular and ,
, Key words: cryo electron microscopy, cryo electron tomography, Crystallography, Super-resolution microscopy, Structural biology, To whom correspondence should be addressed (email: klaholz@igbmc.fr) 2 Present address: European Molecular Biology Laboratory (EMBL), Structural and Computational Biology Unit, vol.1, p.69117
, CMOS, complementary metal oxide semiconductor
, DDD, direct detection device
, DQE, detective quantum efficiency; EcR, ecdysone receptor; FIB, focussed-ion beam; FRISBI, French Infrastructure for Integrated Structural Biology; Instruct, Integrated Structural Biology Infrastructure for Europe; kDa
, MSA, multi-variate statistical analysis; mRNA, messenger RNA; rRNA, ribosomal RNA; SEM, scanning electron microscopy
, SMLM, single-molecule localisation microscopy; tRNA, transfer RNA; 2D, two dimensional
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