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Title: Guiding synchrotron X-ray diffraction by multimodal video-rate protein crystal imaging

Synchronous digitization, in which an optical sensor is probed synchronously with the firing of an ultrafast laser, was integrated into an optical imaging station for macromolecular crystal positioning prior to synchrotron X-ray diffraction. Using the synchronous digitization instrument, second-harmonic generation, two-photon-excited fluorescence and bright field by laser transmittance were all acquired simultaneously with perfect image registry at up to video-rate (15 frames s –1). A simple change in the incident wavelength enabled simultaneous imaging by two-photon-excited ultraviolet fluorescence, one-photon-excited visible fluorescence and laser transmittance. Development of an analytical model for the signal-to-noise enhancement afforded by synchronous digitization suggests a 15.6-fold improvement over previous photon-counting techniques. This improvement in turn allowed acquisition on nearly an order of magnitude more pixels than the preceding generation of instrumentation and reductions of well over an order of magnitude in image acquisition times. These improvements have allowed detection of protein crystals on the order of 1 µm in thickness under cryogenic conditions in the beamline. Lastly, these capabilities are well suited to support serial crystallography of crystals approaching 1 µm or less in dimension.
Authors:
 [1] ;  [1] ;  [1] ;  [1] ;  [2] ;  [1] ;  [2] ;  [1] ;  [1] ;  [1] ;  [2] ;  [1]
  1. Purdue Univ., West Lafayette, IN (United States)
  2. Argonne National Lab. (ANL), Argonne, IL (United States)
Publication Date:
Grant/Contract Number:
AC02-06CH11357
Type:
Accepted Manuscript
Journal Name:
Journal of Synchrotron Radiation (Online)
Additional Journal Information:
Journal Name: Journal of Synchrotron Radiation (Online); Journal Volume: 23; Journal Issue: 4; Journal ID: ISSN 1600-5775
Publisher:
International Union of Crystallography
Research Org:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org:
National Institutes of Health (NIH); USDOE
Country of Publication:
United States
Language:
English
Subject:
73 NUCLEAR PHYSICS AND RADIATION PHYSICS; nonlinear optical microscopy; protein crystal; synchronous digitization
OSTI Identifier:
1339575

Newman, Justin A., Zhang, Shijie, Sullivan, Shane Z., Dow, Ximeng Y., Becker, Michael, Sheedlo, Michael J., Stepanov, Sergey, Carlsen, Mark S., Everly, R. Michael, Das, Chittaranjan, Fischetti, Robert F., and Simpson, Garth J.. Guiding synchrotron X-ray diffraction by multimodal video-rate protein crystal imaging. United States: N. p., Web. doi:10.1107/S1600577516005919.
Newman, Justin A., Zhang, Shijie, Sullivan, Shane Z., Dow, Ximeng Y., Becker, Michael, Sheedlo, Michael J., Stepanov, Sergey, Carlsen, Mark S., Everly, R. Michael, Das, Chittaranjan, Fischetti, Robert F., & Simpson, Garth J.. Guiding synchrotron X-ray diffraction by multimodal video-rate protein crystal imaging. United States. doi:10.1107/S1600577516005919.
Newman, Justin A., Zhang, Shijie, Sullivan, Shane Z., Dow, Ximeng Y., Becker, Michael, Sheedlo, Michael J., Stepanov, Sergey, Carlsen, Mark S., Everly, R. Michael, Das, Chittaranjan, Fischetti, Robert F., and Simpson, Garth J.. 2016. "Guiding synchrotron X-ray diffraction by multimodal video-rate protein crystal imaging". United States. doi:10.1107/S1600577516005919. https://www.osti.gov/servlets/purl/1339575.
@article{osti_1339575,
title = {Guiding synchrotron X-ray diffraction by multimodal video-rate protein crystal imaging},
author = {Newman, Justin A. and Zhang, Shijie and Sullivan, Shane Z. and Dow, Ximeng Y. and Becker, Michael and Sheedlo, Michael J. and Stepanov, Sergey and Carlsen, Mark S. and Everly, R. Michael and Das, Chittaranjan and Fischetti, Robert F. and Simpson, Garth J.},
abstractNote = {Synchronous digitization, in which an optical sensor is probed synchronously with the firing of an ultrafast laser, was integrated into an optical imaging station for macromolecular crystal positioning prior to synchrotron X-ray diffraction. Using the synchronous digitization instrument, second-harmonic generation, two-photon-excited fluorescence and bright field by laser transmittance were all acquired simultaneously with perfect image registry at up to video-rate (15 frames s–1). A simple change in the incident wavelength enabled simultaneous imaging by two-photon-excited ultraviolet fluorescence, one-photon-excited visible fluorescence and laser transmittance. Development of an analytical model for the signal-to-noise enhancement afforded by synchronous digitization suggests a 15.6-fold improvement over previous photon-counting techniques. This improvement in turn allowed acquisition on nearly an order of magnitude more pixels than the preceding generation of instrumentation and reductions of well over an order of magnitude in image acquisition times. These improvements have allowed detection of protein crystals on the order of 1 µm in thickness under cryogenic conditions in the beamline. Lastly, these capabilities are well suited to support serial crystallography of crystals approaching 1 µm or less in dimension.},
doi = {10.1107/S1600577516005919},
journal = {Journal of Synchrotron Radiation (Online)},
number = 4,
volume = 23,
place = {United States},
year = {2016},
month = {5}
}