Transmission electron microscopy (TEM) of rapidly frozen biological specimens, or cryo-EM, would benefit from the development of a phase plate for in-focus phase contrast imaging. Several types of phase plates have been investigated, but rapid electrostatic charging of all such devices has hindered these efforts. Here, we demonstrate electron phase manipulation with a high-intensity continuous-wave laser beam, and use it as a phase plate for TEM. We demonstrate the laser phase plate by imaging an amorphous carbon film. The laser phase plate provides a stable and tunable phase shift without electrostatic charging or unwanted electron scattering. These results suggest the possibility for dose-efficient imaging of unstained biological macromolecules and cells.
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Data and materials availability
The raw TEM images collected over the course of this study are available from the corresponding author upon request.
The code for fitting the light wave micrographs is available from the corresponding author upon request.
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We thank R. Adhikari, B. Buijsse, W. T. Carlisle, A. Chintangal, E. Copenhaver, P. Dona, S. Goobie, P. Grob, B. G. Han, P. Haslinger, M. Jaffe, F. Littlefield, G. W. Long, E. Nogales, Z. Pagel, R. H. Parker, X. Wu, V. Xu and J. Ye for helpful discussions and assistance in various aspects of the experiment. This work was supported by the US National Institutes of Health grant 5 R01 GM126011-02, the US National Science Foundation Grant no. 1040543, the David and Lucile Packard Foundation grant 2009-34712, and Bakar Fellows Program. O.S. is supported by the Human Frontier Science Program postdoctoral fellowship LT000844/2016-C. J.J.A. is supported by the US National Science Foundation Graduate Research Fellowship Program Grant no. D.G.E. 1752814. S.L.C. is supported by the Howard Hughes Medical Institute Hanna H. Gray Fellows Program Grant no. GT11085.
O.S., J.J.A., R.M.G. and H.M. are inventors on US patent application no. 15/939,028. All other authors have no competing interests.
Peer review information Allison Doerr was the primary editor on this article and managed its editorial process and peer review in collaboration with the rest of the editorial team
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Schwartz, O., Axelrod, J.J., Campbell, S.L. et al. Laser phase plate for transmission electron microscopy. Nat Methods 16, 1016–1020 (2019) doi:10.1038/s41592-019-0552-2