Hantke, M. F.Hasse, D.Ekeberg, T.John, K.Svenda, M.Loh, D.Martin, A. V.Timneanu, N.Larsson, D. S. D.Van Der Schot, G.Carlsson, G. H.Ingelman, M.Andreasson, J.Westphal, D.Iwan, B.Uetrecht, C.Bielecki, J.Liang, M.Stellato, F.DePonte, D. P.Bari, S.Hartmann, R.Kimmel, N.Kirian, R. A.Seibert, M. M.Mühlig, K.Schorb, S.Ferguson, K.Bostedt, C.Carron, S.Bozek, J. D.Rolles, DanielRudenko, ArtemFoucar, LutzEpp, S. W.Chapman, H. N.Barty, A.Andersson, I.Hajdu, J.Maia, F. R. N. C.2017-04-102017-04-102016-08-01http://hdl.handle.net/2097/35352Citation: Hantke, M. F., Hasse, D., Ekeberg, T., John, K., Svenda, M., Loh, D., . . . Maia, F. R. N. C. (2016). A data set from flash X-ray imaging of carboxysomes. Scientific Data, 3. doi:10.1038/sdata.2016.61Ultra-intense femtosecond X-ray pulses from X-ray lasers permit structural studies on single particles and biomolecules without crystals. We present a large data set on inherently heterogeneous, polyhedral carboxysome particles. Carboxysomes are cell organelles that vary in size and facilitate up to 40% of Earth's carbon fixation by cyanobacteria and certain proteobacteria. Variation in size hinders crystallization. Carboxysomes appear icosahedral in the electron microscope. A protein shell encapsulates a large number of Rubisco molecules in paracrystalline arrays inside the organelle. We used carboxysomes with a mean diameter of 115±26 nm from Halothiobacillus neapolitanus. A new aerosol sample-injector allowed us to record 70,000 low-noise diffraction patterns in 12 min. Every diffraction pattern is a unique structure measurement and high-throughput imaging allows sampling the space of structural variability. The different structures can be separated and phased directly from the diffraction data and open a way for accurate, high-throughput studies on structures and structural heterogeneity in biology and elsewhere.Attribution 4.0 International (CC BY 4.0)Article