title: Two promising future developments of cryo-EM: capturing short-lived states and mapping a continuum of states of a macromolecule.
abstract: The capabilities and application range of cryogenic electron microscopy (cryo-EM) method have expanded vastly in the last two years, thanks to the advances provided by direct detection devices and computational classification tools. We take this review as an opportunity to sketch out promising developments of cryo-EM in two important directions: (i) imaging of short-lived states (10-1000 ms) of biological molecules by using time-resolved cryo-EM, particularly the mixing-spraying method and (ii) recovering an entire continuum of coexisting states from the same sample by employing a computational technique called manifold embedding. It is tempting to think of combining these two methods, to elucidate the way the states of a molecular machine such as the ribosome branch and unfold. This idea awaits further developments of both methods, particularly by increasing the data yield of the time-resolved cryo-EM method and by developing the manifold embedding technique into a user-friendly workbench.
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Columbia University, 650 W. 168th Street, New York, NY 10032, USA.
authors.forename: Bo
authors.initials: B
authors.lastname: Chen
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Columbia University, 650 W. 168th Street, New York, NY 10032, USA Department of Biological Sciences, Columbia University, New York, NY 10027, USA Howard Hughes Medical Institute, Columbia University, New York, NY 10032, USA jf2192@cumc.columbia.edu.
authors.forename: Joachim
authors.initials: J
authors.lastname: Frank
mesh_terms: D020285:Cryoelectron Microscopy / Q000379:methods*; D046911:Macromolecular Substances / Q000032:analysis*; D011487:Protein Conformation; D012270:Ribosomes / Q000737:chemistry; D013995:Time
chemical_list: D046911:Macromolecular Substances
keywords: classification; manifold embedding; microfluidics; ribosome; time-resolved imaging; translation