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Title: Electrospray sample injection for single-particle imaging with x-ray lasers

Abstract

The possibility of imaging single proteins constitutes an exciting challenge for x-ray lasers. Despite encouraging results on large particles, imaging small particles has proven to be difficult for two reasons: not quite high enough pulse intensity from currently available x-ray lasers and, as we demonstrate here, contamination of the aerosolized molecules by nonvolatile contaminants in the solution. The amount of contamination on the sample depends on the initial droplet size during aerosolization. Here, we show that, with our electrospray injector, we can decrease the size of aerosol droplets and demonstrate virtually contaminant-free sample delivery of organelles, small virions, and proteins. The results presented here, together with the increased performance of next-generation x-ray lasers, constitute an important stepping stone toward the ultimate goal of protein structure determination from imaging at room temperature and high temporal resolution.

Authors:
ORCiD logo [1]; ORCiD logo [2]; ORCiD logo [3];  [4]; ORCiD logo [4]; ORCiD logo [4]; ORCiD logo [4];  [5]; ORCiD logo [4]; ORCiD logo [5];  [6]; ORCiD logo [4]; ORCiD logo [4];  [4]; ORCiD logo [4];  [4]; ORCiD logo [4];  [4];  [7]; ORCiD logo [8] more »; ORCiD logo [9]; ORCiD logo [10];  [11];  [12];  [11]; ORCiD logo [13];  [4];  [11];  [11]; ORCiD logo [11]; ORCiD logo [14];  [15];  [11];  [16];  [11]; ORCiD logo [4]; ORCiD logo [17]; ORCiD logo [18]; ORCiD logo [19] « less
  1. Uppsala Univ., Uppsala (Sweden); European XFEL GmbH, Schenefeld (Germany)
  2. Uppsala Univ., Uppsala (Sweden); Oxford Univ., Oxford (United Kingdom); Refeyn Ltd., Oxford (United Kingdom)
  3. Uppsala Univ., Uppsala (Sweden); National Univ. of Singapore (Singapore)
  4. Uppsala Univ., Uppsala (Sweden)
  5. Uppsala Univ., Uppsala (Sweden); KTH Royal Institute of Technology, Stockholm (Sweden)
  6. La Trobe Univ., Melbourne, VIC (Australia)
  7. Academy of Sciences of the Czech Republic, Prague (Czech Republic)
  8. Osaka Univ., Osaka (Japan); Hiroshima Univ., Hiroshima (Japan)
  9. Uppsala Univ., Uppsala (Sweden); Utrecht Univ., Utrecht (Netherlands)
  10. National Univ. of Singapore (Singapore)
  11. SLAC National Accelerator Lab., Menlo Park, CA (United States)
  12. Argonne National Lab. (ANL), Argonne, IL (United States); Northwestern Univ., Evanston, IL (United States)
  13. SLAC National Accelerator Lab., Menlo Park, CA (United States); Leibniz Univ. Hannover, Hannover (Germany)
  14. Argonne National Lab. (ANL), Argonne, IL (United States)
  15. Technische Univ. Berlin, Berlin (Germany)
  16. SLAC National Accelerator Lab., Menlo Park, CA (United States); Brookhaven National Lab. (BNL), Upton, NY (United States)
  17. Academy of Sciences of the Czech Republic, Prague (Czech Republic); Chalmers Univ. of Technology, Gothenburg (Sweden)
  18. Uppsala Univ., Uppsala (Sweden); Academy of Sciences of the Czech Republic; Prague (Czech Republic)
  19. Uppsala Univ., Uppsala (Sweden); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States); Argonne National Lab. (ANL), Argonne, IL (United States); Brookhaven National Lab. (BNL), Upton, NY (United States)
Sponsoring Org.:
Swedish Foundation for Strategic Research (SSF); USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22). Chemical Sciences, Geosciences & Biosciences Division; Swedish Research Council (SRC); Knut and Alice Wallenberg Foundation; European Research Council (ERC); European Regional Development Fund (ERDF); Wellcome Trust; Volkswagen Foundation; USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1532400
Alternate Identifier(s):
OSTI ID: 1559029; OSTI ID: 1571429
Report Number(s):
BNL-212202-2019-JAAM
Journal ID: ISSN 2375-2548
Grant/Contract Number:  
AC02-76SF00515; AC02-06CH11357; SC0012704
Resource Type:
Accepted Manuscript
Journal Name:
Science Advances
Additional Journal Information:
Journal Volume: 5; Journal Issue: 5; Journal ID: ISSN 2375-2548
Publisher:
AAAS
Country of Publication:
United States
Language:
English
Subject:
47 OTHER INSTRUMENTATION; 46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND TECHNOLOGY

Citation Formats

Bielecki, Johan, Hantke, Max F., Daurer, Benedikt J., Reddy, Hemanth K. N., Hasse, Dirk, Larsson, Daniel S. D., Gunn, Laura H., Svenda, Martin, Munke, Anna, Sellberg, Jonas A., Flueckiger, Leonie, Pietrini, Alberto, Nettelblad, Carl, Lundholm, Ida, Carlsson, Gunilla, Okamoto, Kenta, Timneanu, Nicusor, Westphal, Daniel, Kulyk, Olena, Higashiura, Akifumi, van der Schot, Gijs, Loh, Ne-Te Duane, Wysong, Taylor E., Bostedt, Christoph, Gorkhover, Tais, Iwan, Bianca, Seibert, M. Marvin, Osipov, Timur, Walter, Peter, Hart, Philip, Bucher, Maximilian, Ulmer, Anatoli, Ray, Dipanwita, Carini, Gabriella, Ferguson, Ken R., Andersson, Inger, Andreasson, Jakob, Hajdu, Janos, and Maia, Filipe R. N. C. Electrospray sample injection for single-particle imaging with x-ray lasers. United States: N. p., 2019. Web. doi:10.1126/sciadv.aav8801.
Bielecki, Johan, Hantke, Max F., Daurer, Benedikt J., Reddy, Hemanth K. N., Hasse, Dirk, Larsson, Daniel S. D., Gunn, Laura H., Svenda, Martin, Munke, Anna, Sellberg, Jonas A., Flueckiger, Leonie, Pietrini, Alberto, Nettelblad, Carl, Lundholm, Ida, Carlsson, Gunilla, Okamoto, Kenta, Timneanu, Nicusor, Westphal, Daniel, Kulyk, Olena, Higashiura, Akifumi, van der Schot, Gijs, Loh, Ne-Te Duane, Wysong, Taylor E., Bostedt, Christoph, Gorkhover, Tais, Iwan, Bianca, Seibert, M. Marvin, Osipov, Timur, Walter, Peter, Hart, Philip, Bucher, Maximilian, Ulmer, Anatoli, Ray, Dipanwita, Carini, Gabriella, Ferguson, Ken R., Andersson, Inger, Andreasson, Jakob, Hajdu, Janos, & Maia, Filipe R. N. C. Electrospray sample injection for single-particle imaging with x-ray lasers. United States. doi:10.1126/sciadv.aav8801.
Bielecki, Johan, Hantke, Max F., Daurer, Benedikt J., Reddy, Hemanth K. N., Hasse, Dirk, Larsson, Daniel S. D., Gunn, Laura H., Svenda, Martin, Munke, Anna, Sellberg, Jonas A., Flueckiger, Leonie, Pietrini, Alberto, Nettelblad, Carl, Lundholm, Ida, Carlsson, Gunilla, Okamoto, Kenta, Timneanu, Nicusor, Westphal, Daniel, Kulyk, Olena, Higashiura, Akifumi, van der Schot, Gijs, Loh, Ne-Te Duane, Wysong, Taylor E., Bostedt, Christoph, Gorkhover, Tais, Iwan, Bianca, Seibert, M. Marvin, Osipov, Timur, Walter, Peter, Hart, Philip, Bucher, Maximilian, Ulmer, Anatoli, Ray, Dipanwita, Carini, Gabriella, Ferguson, Ken R., Andersson, Inger, Andreasson, Jakob, Hajdu, Janos, and Maia, Filipe R. N. C. Fri . "Electrospray sample injection for single-particle imaging with x-ray lasers". United States. doi:10.1126/sciadv.aav8801. https://www.osti.gov/servlets/purl/1532400.
@article{osti_1532400,
title = {Electrospray sample injection for single-particle imaging with x-ray lasers},
author = {Bielecki, Johan and Hantke, Max F. and Daurer, Benedikt J. and Reddy, Hemanth K. N. and Hasse, Dirk and Larsson, Daniel S. D. and Gunn, Laura H. and Svenda, Martin and Munke, Anna and Sellberg, Jonas A. and Flueckiger, Leonie and Pietrini, Alberto and Nettelblad, Carl and Lundholm, Ida and Carlsson, Gunilla and Okamoto, Kenta and Timneanu, Nicusor and Westphal, Daniel and Kulyk, Olena and Higashiura, Akifumi and van der Schot, Gijs and Loh, Ne-Te Duane and Wysong, Taylor E. and Bostedt, Christoph and Gorkhover, Tais and Iwan, Bianca and Seibert, M. Marvin and Osipov, Timur and Walter, Peter and Hart, Philip and Bucher, Maximilian and Ulmer, Anatoli and Ray, Dipanwita and Carini, Gabriella and Ferguson, Ken R. and Andersson, Inger and Andreasson, Jakob and Hajdu, Janos and Maia, Filipe R. N. C.},
abstractNote = {The possibility of imaging single proteins constitutes an exciting challenge for x-ray lasers. Despite encouraging results on large particles, imaging small particles has proven to be difficult for two reasons: not quite high enough pulse intensity from currently available x-ray lasers and, as we demonstrate here, contamination of the aerosolized molecules by nonvolatile contaminants in the solution. The amount of contamination on the sample depends on the initial droplet size during aerosolization. Here, we show that, with our electrospray injector, we can decrease the size of aerosol droplets and demonstrate virtually contaminant-free sample delivery of organelles, small virions, and proteins. The results presented here, together with the increased performance of next-generation x-ray lasers, constitute an important stepping stone toward the ultimate goal of protein structure determination from imaging at room temperature and high temporal resolution.},
doi = {10.1126/sciadv.aav8801},
journal = {Science Advances},
number = 5,
volume = 5,
place = {United States},
year = {2019},
month = {5}
}

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Figures / Tables:

Figure 1 Figure 1: ES aerosol injector. (A) Design of the ES aerosol injector. In the aerosolization chamber, the ES nebulizer generates droplets that are neutralized with a 210Po alpha emitter. The ES nebulizer is operated in an atmosphere of N2 and CO2 at 1 bar. The aerosol is transported through twomore » nozzle-skimmer assemblies, where excess gas is pumped away. At a reduced pressure of 1 to 10 mbar, the aerosol enters the aerosol lens stack, which focuses it to a narrow particle beam entering the experimental chamber, which is held at a pressure of 10−6 to 10−5 mbar to match requirements for XFEL imaging. (B) Size distributions of initial droplets for ES (green) and GDVN (blue) aerosols determined by RSM (top) and XFEL diffraction (bottom). The results of the two sizing methods are comparable within the limits of reproducibility expected for the manually manufactured nozzles and variations in operational parameters, such as pressures, voltage, and flow rate. (C) RSM size distributions of aerosolized particles from carboxysome sample (purple) and from its buffer solution (red). Data collected on electrosprayed particles are shown in the first panel (median, 95 nm; FWHM, 14 nm), and data collected on particles injected by GDVN at two different pressure configurations (Table 2) are shown in the second (median, 102 nm; FWHM, 17 nm) and third panels (median, 105 nm; FWHM, 17 nm). Dashed lines indicate the detection limit.« less

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