research papers 188 doi:10.1107/S1600576713029944 J. Appl. Cryst. (2014). 47, 188–197 Journal of Applied Crystallography ISSN 1600-5767 Received 25 June 2013 Accepted 1 November 2013 # 2014 International Union of Crystallography Multiple application X-ray imaging chamber for single-shot diffraction experiments with femtosecond X-ray laser pulses Changyong Song, a * Kensuke Tono, b Jaehyun Park, a Tomio Ebisu, a Sunam Kim, a Hiroki Shimada, a Sangsoo Kim, a Marcus Gallagher-Jones, a,c Daewoong Nam, a,d Takahiro Sato, a Tadashi Togashi, b Kanade Ogawa, a Yasumasa Joti, b Takashi Kameshima, b Shun Ono, a Takaki Hatsui, a So Iwata, a,e,f,g Makina Yabashi a and Tetsuya Ishikawa a a RIKEN SPring-8 Center, 1-1-1 Kouto, Sayo, Hyogo 679-5148, Japan, b Japan Synchrotron Radiation Research Institute, 1-1-1 Kouto, Sayo, Hyogo 679-5198, Japan, c Institute of Integrative Biology, University of Liverpool, Liverpool L69 7ZB, England, d Department of Physics, Pohang University of Science and Technology, Pohang 790-784, Republic of Korea, e Department of Cell Biology, Graduate School of Medicine, Kyoto University, Yoshidakonoe-cho, Sakyo-ku, Kyoto, 606-8501, Japan, f Membrane Protein Laboratory, Imperial College London, Diamond House, Harwell Science and Innovation Campus, Didcot, Oxfordshire OX11 0DE, England, and g Membrane Protein Crystallography Project, Research Acceleration Program, JST, Yoshidakonoe-cho, Sakyo-ku, Kyoto, 606-8501, Japan. Correspondence e-mail: cysong@spring8.or.jp X-ray free-electron lasers (XFELs) provide intense (10 12 photons per pulse) coherent X-rays with ultra-short (10 14 s) pulse lengths. X-rays of such an unprecedented nature have introduced new means of atomic scale structural investigations, and discoveries are still ongoing. Effective use of XFELs would be further accelerated on a highly adaptable platform where most of the new experiments can be realized. Introduced here is the multiple-application X-ray imaging chamber (MAXIC), which is able to carry out various single-pulse diffraction experiments including single-shot imaging, nanocrystallographic data acquisition and ultra-fast pump–probe scattering for specimens in solid, liquid and gas phases. The MAXIC established at the SPring-8 a ˚ ngstro ¨ m compact free- electron laser (SACLA) has demonstrated successful applications in the aforementioned experiments, but is not limited to them. Also introduced are recent experiments on single-shot diffraction imaging of Au nanoparticles and serial crystallographic data collection of lysozyme crystals at SACLA. 1. Introduction The insight of Max von Laue in 1912 to propose the diffraction of X-rays by regularly arranged atoms in crystals gave birth to X-ray crystallography. Since then, the invention of new X-ray measurement techniques has been incessant, making it one of the most reliable structural probes at the atomic scale (Hendrickson, 2013). The desire to acquire signals from ever smaller samples, to distinguish subtle structural differences, to unveil the structures of noncrystalline specimens, to observe transient features etc. has prompted the new development of advanced X-ray sources. The advent of the X-ray free-electron laser (XFEL) provides new X-rays of an unprecedented nature allowing greater innovation. XFELs provide femtosecond (fs) X-ray pulses containing 10 12 photons of near-complete spatial coherence (Acker- mann et al., 2007; Ishikawa et al., 2012; Vartanyants et al. , 2011). These intense coherent X-rays enable us to acquire diffraction patterns from small crystals or macromolecular complexes of sizes ranging from submicrometre scales to several micrometres (Chapman et al., 2011; Seibert et al., 2011; Boutet et al., 2012). The intense radiation damages samples irreversibly, but the X-rays in a femtosecond pulse escape the samples before the damage process starts to appear (Neutze et al., 2000; Gaffney & Chapman, 2007; Park et al., 2012; Barty et al., 2012). This enables the recording of snapshot diffraction patterns from samples in an almost damage-free condition, insofar as new samples are supplied for every new X-ray pulse. Supported by the unique characteristics of XFELs, the past couple of years have seen the introduction of various new experiments such as serial femtosecond protein crystal- lography, single-shot diffraction imaging etc., which could significantly reduce the limitation of preparing sizeable protein crystals, or make an important step toward single molecular imaging (Miao et al. , 2001; Barty et al. , 2013). The emergence of these new measurement techniques and scientific discoveries has stimulated strong interest in XFELs (Kern et al., 2012; Loh et al., 2012; Koopmann et al., 2012; Starodub et al. , 2012; Schlichting & Miao, 2012; Wochner et al.,