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

Science advances (2019-05-07)
Johan Bielecki, Max F Hantke, Benedikt J Daurer, Hemanth K N Reddy, Dirk Hasse, Daniel S D Larsson, Laura H Gunn, Martin Svenda, Anna Munke, Jonas A Sellberg, Leonie Flueckiger, Alberto Pietrini, Carl Nettelblad, Ida Lundholm, Gunilla Carlsson, Kenta Okamoto, Nicusor Timneanu, Daniel Westphal, Olena Kulyk, Akifumi Higashiura, Gijs van der Schot, Ne-Te Duane Loh, Taylor E Wysong, Christoph Bostedt, Tais Gorkhover, Bianca Iwan, M Marvin Seibert, Timur Osipov, Peter Walter, Philip Hart, Maximilian Bucher, Anatoli Ulmer, Dipanwita Ray, Gabriella Carini, Ken R Ferguson, Inger Andersson, Jakob Andreasson, Janos Hajdu, Filipe R N C Maia

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.