John Morse

High-resolution protein structure determination by serial femtosecond crystallography

Size Matters Less X-ray crystallography is a central research tool for uncovering the structures of proteins and other macromolecules. However, its applicability typically requires growth of large crystals, in part because a sufficient number of molecules must be present in the lattice for the sample to withstand x-ray—induced damage. Boutet et al. (p. 362, published online 31 May) now demonstrate that the intense x-ray pulses emitted by a free-electron laser source can yield data in few enough exposures to uncover the high-resolution structure of microcrystals. A powerful x-ray laser source can probe proteins in detail using much smaller crystals than previously required. Structure determination of proteins and other macromolecules has historically required the growth of high-quality crystals sufficiently large to diffract x-rays efficiently while withstanding radiation damage. We applied serial femtosecond crystallography (SFX) using an x-ray free-electron laser (XFEL) to obtain high-resolution structural information from microcrystals (less than 1 micrometer by 1 micrometer by 3 micrometers) of the well-characterized model protein lysozyme. The agreement with synchrotron data demonstrates the immediate relevance of SFX for analyzing the structure of the large group of difficult-to-crystallize molecules.

The CSPAD megapixel x-ray camera at LCLS

The Linear Coherent Light Source (LCLS), a free electron laser operating from 250eV to10keV at 120Hz, is opening windows on new science in biology, chemistry, and solid state, atomic, and plasma physics1,2. The FEL provides coherent x-rays in femtosecond pulses of unprecedented intensity. This allows the study of materials on up to 3 orders of magnitude shorter time scales than previously possible. Many experiments at the LCLS require a detector that can image scattered x-rays on a per-shot basis with high efficiency and excellent spatial resolution over a large solid angle and both good S/N (for single-photon counting) and large dynamic range (required for the new coherent x-ray diffractive imaging technique3). The Cornell-SLAC Pixel Array Detector (CSPAD) has been developed to meet these requirements. SLAC has built, characterized, and installed three full camera systems at the CXI and XPP hutches at LCLS. This paper describes the camera system and its characterization and performance.

ID16B: a hard X-ray nanoprobe beamline at the ESRF for nano-analysis.

ID16B is a versatile hard X-ray nanoprobe devoted to X-ray nano-analysis. It combines X-ray fluorescence, X-ray diffraction, X-ray absorption spectroscopy and 2D/3D X-ray imaging techniques.

Wavelength-dispersive spectrometer for X-ray microfluorescence analysis at the X-ray microscopy beamline ID21 (ESRF)

A polycapillary-based wavelength-dispersive spectrometer is reported. The design consideration as well as operation characteristics are presented.

Large-acceptance diamond planar refractive lenses manufactured by laser cutting

For the first time, single-crystal diamond planar refractive lenses have been fabricated by laser micromachining in 300 µm-thick diamond plates which were grown by chemical vapour deposition. Linear lenses with apertures up to 1 mm and parabola apex radii up to 500 µm were manufactured and tested at the ESRF ID06 beamline. The large acceptance of these lenses allows them to be used as beam-conditioning elements. Owing to the unsurpassed thermal properties of single-crystal diamond, these lenses should be suitable to withstand the extreme flux densities expected at the planned fourth-generation X-ray sources.

The Cornell-SLAC pixel array detector at LCLS

The Cornell-SLAC pixel array detector (CSpad) is a general-purpose integrating hybrid pixel x-ray camera developed for use at the Linear Coherent Light Source (LCLS) x-ray free electron laser at the SLAC National Accelerator Laboratory (SLAC). The detector has a full well capacity of about 2.Sk photons in low-gain mode and a SIN of about 6 in high-gain mode. Its 2.3M pixels are read out at 120 Hz. The detector comprises 32 500μm silicon sensors bump-bonded to 64 185×194-pixel ASICs. The pixel size is 110μm. The water-cooled detector quadrants can be radially moved in-situ to vary the beam aperture. SLAC has built, calibrated, and optimized three complete camera systems based on a sensor and ASIC designed by Cornell. The camera is read out by a DAQ system which provides extensive online monitoring and prompt analysis capabilities. We have also built a dozen smaller cameras in a portable form-factor for use in confined spaces and for ease of development, testing, and deployment. Through 2012 user experiments have taken almost a petabyte of data with these detectors in a variety of applications. We have extensively tested the detector at synchrotrons and with an x-ray tube, in addition to commissioning tests at the LCLS, investigating linearity, cross-talk, homogeneity, and radiation hardness. The SLAC detector group is deploying improved support infrastructure and an updated ASIC and electronics based on this experience. This paper describes the instrument, its calibration and performance, and presents preliminary results from the updated camera.

An upgrade beamline for combined wide, small and ultra small-angle x-ray scattering at the ESRF

This contribution presents the main design features of the upgraded beamline ID02 (TRUSAXS). The beamline combines different small-angle X-ray scattering techniques in one unique instrument. The key component of this instrument is an evacuated (5×10−3 mbar) stainless steel detector tube of length 34 m and diameter 2 m. Three different detectors (Rayonix MX170, Pilatus 300K and FReLoN 4M) are housed inside a motorized wagon which travels along a rail system with very low parasitic lateral movements (± 0.3 mm). This system allows automatically changing the sample-to-detector distance from about 1 m to 31 m and selecting the desired detector. In addition, a wide angle detector (Rayonix LX170) is installed just above the entrance cone of the tube for optional wide-angle X-ray scattering measurements. The beamstop system enables monitoring of the X-ray beam intensity in addition to blocking the primary beam, and automated insertion of selected masks behind the primary beamstop. The focusing optics and collimat...

MHz frame rate hard X-ray phase-contrast imaging using synchrotron radiation

Third generation synchrotron light sources offer high photon flux, partial spatial coherence, and ~10-10 s pulse widths. These enable hard X-ray phase-contrast imaging (XPCI) with single-bunch temporal resolutions. In this work, we exploited the MHz repetition rates of synchrotron X-ray pulses combined with indirect X-ray detection to demonstrate the potential of XPCI with millions of frames per second multiple-frame recording. This allows for the visualization of aperiodic or stochastic transient processes which are impossible to be realized using single-shot or stroboscopic XPCI. We present observations of various phenomena, such as crack tip propagation in glass, shock wave propagation in water and explosion during electric arc ignition, which evolve in the order of km/s (µm/ns).

3DX: a micromachined silicon crystallographic x-ray detector

We are developing pixel detectors for macromolecular crystallography, in which diffracted X-rays are directly absorbed by high-resistivity, crystalline silicon that has been micro-machined by inductively-coupled plasma etching. Arrays of 64 × 64 holes at 150 μm pitch are first formed by etching through the entire silicon bulk, then backfilled with polysilicon that is doped to create conducting p and n type columnar electrodes. When reverse biased, these electrodes generate electric fields that define the individual pixels. By forming conducting polysilicon on the sides of the sensors, which are cut-out of the silicon wafer by plasma etching, the entire surface of the detector may be made active. CMOS readout integrated circuits are conductively bump bonded behind each 3D detector, providing a direct connection to every pixel. A large array will be assembled with no insensitive bands along the edges by overlapping these sensors, each of area 0.96cm2. This detector will measure X-ray signal intensities of up to 105 events/pixel/sec without any pile-up loss, by using an integration method that retains the benefits of discrete photon counting. The detector sensitivity will be highly uniform, it will not exhibit any dark signal or spurious noise, and no geometric distortion will occur within each sensor.

Energy and position resolution of germanium microstrip detectors at X-ray energies from 15 to 100 keV

In addition to their far greater X-ray detection efficiency, germanium strip detectors offer superior energy and position resolution as compared to those fabricated of silicon for energies in the range of 15 to 100 keV. We have characterized 200-/spl mu/m strip pitch detectors fabricated by two different processes. By scanning a 10-/spl mu/m-wide monochromatic synchrotron X-ray beam across these detectors, measurements were made on both spectral energy response and spatial resolution. X-rays absorbed between neighboring diode strips suffer from charge diffusion splitting of their signals which seriously degrades the detector performance, but by reconstructing events using an energy-sum coincidence algorithm we succeeded in producing artifact-free spectra with energy resolution 1000, and count uniformities across the detector surface <1.5% for energies below 60 keV. The experimentally measured energy spectra show remarkable agreement with those predicted by computer simulation, in which the EGS4 code for photon absorption is combined with a simple algorithm to account for charge diffusion.