High-throughput data collection for macromolecular crystallography requires an automated sample mounting and alignment system for cryo-protected crystals that functions reliably when integrated into protein-crystallography beamlines at synchrotrons. Rapid mounting and dismounting of the samples increases the efficiency of the crystal screening and data collection processes, where many crystals can be tested for the quality of diffraction. The sample-mounting subsystem has random access to 112 samples, stored under liquid nitrogen. Results of extensive tests regarding the performance and reliability of the system are presented. To further increase throughput, we have also developed a sample transport/storage system based on "puck-shaped" cassettes, which can hold sixteen samples each. Seven cassettes fit into a standard dry shipping Dewar. The capabilities of a robotic crystal mounting and alignment system with instrumentation control software and a relational database allows for automated screening and data collection to be developed.

SynopsisThree near identical protein crystallography beamlines with a single 6 Tesla peak field superconducting dipole bend magnet as the source have been built at the 1.9 GeV Advanced Light Source. The design and performance of this new facility is described. AbstractAt the Advanced Light Source (ALS), three protein crystallography (PX) beamlines have been built that use as a source one of the three 6 Tesla single pole superconducting bending magnets (superbends) that were recently installed in the ring. The use of such single pole superconducting bend magnets enables the development of a hard x-ray program on a relatively low energy 1.9 GeV ring without taking up insertion device straight sections. The source is of relatively low power, but due to the small electron beam emittance, it has high brightness. X-ray optics are required to preserve the brightness and to match the illumination requirements for protein crystallography. This was achieved by means of a collimating premirror bent to a plane parabola, a double crystal monochromator followed by a toroidal mirror that focuses in the horizontal direction with a 2:1 demagnification. This optical arrangement partially balances aberrations from the collimating and toroidal mirrors such that a tight focused spot size is achieved. The optical properties of the beamline are an excellent match to those required by the small protein crystals that are typically measured. The design and performance of these new beamlines are described.

The Advanced Photoinjector Experiment (APEX) at the Lawrence Berkeley National Laboratory is dedicated to the development of a high-brightness high-repetition rate (MHz-class) electron injector for x-ray free-electron laser (FEL) and other applications where high repetition rates and high brightness are simultaneously required. The injector is based on a new concept rf gun utilizing a normal-conducting (NC) cavity resonating in the VHF band at 186 MHz, and operating in continuous wave (cw) mode in conjunction with high quantum efficiency photocathodes capable of delivering the required charge at MHz repetition rates with available laser technology. The APEX activities are staged in three phases. In phase 0, the NC cw gun is built and tested to demonstrate the major milestones to validate the gun design and performance. Also, starting in phase 0 and continuing in phase I, different photocathodes are tested at the gun energy and at full repetition rate for validating candidate materials to operate in a high-repetition rate FEL. In phase II, a room-temperature pulsed linac is added for accelerating the beam at several tens of MeV to reduce space charge effects and allow the measurement of the brightness of the beam from the gun when integrated in an injector scheme. The installation of the phase 0 beam line and the commissioning of the VHF gun are completed, phase I components are under fabrication, and initial design and specification of components and layout for phase II are under way. This paper presents the phase 0 commissioning results with emphasis on the experimental milestones that have successfully demonstrated the APEX gun capability of operating at the required performance.

A flat, relatively thin (9 mm) xenon-filled multiwire proportional counter with two-dimensional, 2/~s delay line readout of a 270 x 300 mm active area has been developed for use as a position-sensitive area X-ray detector in the 8 keV energy region (Cu Ka) used in crystallographic structure work with large biological molecules. Its quantum detection efficiency for 8 keV Xray photons is about 0.5, a value which is spatially uniform to within _+ 2%. Its dead-time loss fraction at a typical data collection rate of 30 000 photons s-1 is 12%. The detector has spatial resolution for X-rays of 0.6 mm FWHM in the horizontal direction and 2 ram, the anode wire spacing, in the vertical direction. The effects of parallax are found to be limited and do not seriously increase the apparent size of the diffracted beams. The position sensitivity of this detector is geometrically linear to within 0-5 mm across its active surface. Routine maintenance of the detector requires the attention of a skilled technician but is not time consuming. For four years, this detector has been used to measure millions of reflection intensities from crystals of many different proteins. The down time due to the detector has averaged less than four days per year, considerably less than the down time of other components of the data collection system. Four new protein structures have now been solved using data from this detector. Also, a considerable amount of data have been collected at higher resolution or at different temperatures with crystals of other proteins.

A multiwire proportional chamber (30 × 30 cm) together with its electronic readout into a large core memory (mass core) has been used successfully as a digital area detector for protein crystallography. The diffraction pattern stored in the mass core can be displayed on a TV monitor. An IBM 1800 computer has fast random access to the mass core and is used on line to estimate the integrated reflection intensities. To characterize this new area detector, the geometric linearity, the resolution and the quantum detection uniformity have been measured. Preliminary results show that with this new system one can collect intensity data from protein crystals about an order of magnitude faster than with the standard diffractometer.