Serial femtosecond crystallography (SFX) provides opportunities to observe the dynamics of macromolecules without causing radiation damage at room temperature. Although SFX provides a biologically more reliable crystal structure than provided by the existing synchrotron sources, there are limitations due to the consumption of many crystal samples. A viscous medium as a carrier matrix reduces the flow rate of the crystal sample from the injector, thereby dramatically reducing sample consumption. However, the currently available media cannot be applied to specific crystal samples owing to reactions between the viscous medium and crystal sample. The discovery and characterisation of a new delivery medium for SFX can further expand its use. Herein, we report the preparation of a polyacrylamide (PAM) injection matrix to determine the crystal structure with an X-ray free-electron laser. We obtained 11,936 and 22,213 indexed images using 0.5 mg lysozyme and 1.0 mg thermolysin, respectively. We determined the crystal structures of lysozyme and thermolysin delivered in PAM at 1.7 Å and 1.8 Å resolutions. The maximum background scattering from PAM was lower than monoolein, a commonly used viscous medium. Our results show that PAM can be used as a sample delivery media in SFX studies.

Bluetooth Low Energy (BLE) is a short-range wireless communication technology aiming at low-cost and low-power communication. The performance evaluation of classical Bluetooth device discovery have been intensively studied using analytical modeling and simulative methods, but these techniques are not applicable to BLE, since BLE has a fundamental change in the design of the discovery mechanism, including the usage of three advertising channels. Recently, there several works have analyzed the topic of BLE device discovery, but these studies are still far from thorough. It is thus necessary to develop a new, accurate model for the BLE discovery process. In particular, the wide range settings of the parameters introduce lots of potential for BLE devices to customize their discovery performance. This motivates our study of modeling the BLE discovery process and performing intensive simulation. This paper is focused on building an analytical model to investigate the discovery probability, as well as the expected discovery latency, which are then validated via extensive experiments. Our analysis considers both continuous and discontinuous scanning modes. We analyze the sensitivity of these performance metrics to parameter settings to quantitatively examine to what extent parameters influence the performance metric of the discovery processes.

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We demonstrate a hard-X-ray self-seeded (HXRSS) free-electron laser (FEL) at Pohang Accelerator Laboratory with an unprecedented peak brightness (3.2 × 10 35 photons/(s•mm 2 •mrad 2 •0.1%BW)). The self-seeded FEL generates hard X-ray pulses with improved spectral purity; the average pulse energy was 0.85 mJ at 9.7 keV, almost as high as in SASE mode; the bandwidth (0.19 eV) is about 1/70 as wide, the peak spectral brightness is 40 times higher than in self-ampli ed spontaneous emission (SASE) mode, and the stability is excellent with > 94% of shots exceeding the average SASE intensity. Using this self-seeded XFEL, we conducted serial femtosecond crystallography (SFX) experiments to map the structure of lysozyme protein; data-quality metrics such as R split , multiplicity, and signal-to-noise ratio for the SFX were substantially increased. We precisely map out the structure of lysozyme protein with substantially better statistics for the diffraction data and signi cantly sharper electron density maps compared to maps obtained using SASE mode.

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Microcrystal delivery methods are pivotal in the use of serial femtosecond crystallography (SFX) to resolve the macromolecular structures of proteins. Here, the development of a novel technique and instruments for efficiently delivering microcrystals for SFX are presented. The new method, which relies on a one‐dimensional fixed‐target system that includes a microcrystal container, consumes an extremely low amount of sample compared with conventional two‐dimensional fixed‐target techniques at ambient temperature. This novel system can deliver soluble microcrystals without highly viscous carrier media and, moreover, can be used as a microcrystal growth device for SFX. Diffraction data collection utilizing this advanced technique along with a real‐time visual servo scan system has been successfully demonstrated for the structure determination of proteinase K microcrystals at 1.85 Å resolution.

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