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Two promising future developments of cryo-EM: capturing short-lived states and mapping a continuum of states of a macromolecule.
Bo Chen and Joachim Frank

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: (1) imaging of short-lived states (10 ms – 1000 ms) of biological molecules by using time-resolved cryo-EM, particularly the mixing-spraying method, and (2) 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.

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