Towards User Operations of SONICC at GM/CA@APS

Second order nonlinear optical imaging of chiral crystals (SONICC), based on femtosecond laser scanning microscopy, has been implemented at GM/CA@APS undulator beamline 23ID-B for rapid protein crystal localization and centering. The technique is based on infrared laser light impinging on non-centrosymmetric crystals of proteins, which selectively may yield a frequency-doubled, visible signal generated by the anharmonic response of the electron cloud of the protein in response to the laser field. One aim of this method is to locate small crystals grown in opaque crystallization media for centering in X-ray beams of only a few microns or less in cross-section [1]. The optical system implemented at the beamline includes `trans' and `epi' detection of Second Harmonic Generation (SHG) signals [2]. In addition, scanning visible laser light across the sample and detecting two-photon excited UV fluorescence (TPE-UVF) provides complementary contrast based on the native fluorescence of proteins. An update on progress towards offering a user-friendly system to users will be provided. Different factors that influence imaging signals and the practice of successfully locating and accurately positioning a crystal via SONICC will be discussed.

High Brightness Beam for Microcrystallography at GM/CA@APS

GM/CA is a world leader in the development of microcrystallography capabilities for biological macromolecules. The combination of the GM/CA-developed quad-mini-beam collimator and advanced rastering and vector collect software tools have revolutionized microcrystallography. Recently, beamline 23ID-B was reconfigured by shifting the focusing optics 3.8 m downstream. This significantly increased the source demagnification and led to a 4-fold increase in the 5- and 10-micron beam intensity. Beamline 23ID-D is also being upgraded. A Pilatus3-6M with a 1.0 mm thick X-ray sensor was commissioned in January 2014 allowing shutterless data collection with high S/N. The detector specifications include 100 Hz frame rate, 10 MHz/pixel count rate, and high X-ray efficiency. The beamline optics and endstation are also being upgraded to provide a high intensity beam whose size can be variable rapidly in the range of 1 - 20 micron, a new air bearing goniometer with a sphere-of-confusion (SOC) of ~100 nm, a miniature sample XYZ stage that allows centering and scanning of a micron-sized crystal, and a new on-axis-visualization system that provides high resolution optical images of sample crystals. Plans are being developed to upgrade the Advance Photon Source storage ring with a Multi-Bend Achromat lattice. The source properties will be dramatically improved primarily by reducing the horizontal source size to be comparable to the vertical source size, resulting in a 2-3 orders of magnitude increase in source brightness. Both beamlines will be significantly improved by the source upgrade. Moreover the new microfocusing optics for 23ID-D will fully exploit the new source and could deliver a 500 nm (FWHM) beam with >2e13 photons/sec. This unprecedented flux density will provide new opportunities and challenges, and allow the study some of the most important problems in biology. Details of these developments will be presented.

Remote Access Capabilities at the GM/CA Beamlines at the APS

The GM/CA facility consists of two undulator source beamlines and a bending magnet beamline at the Advanced Photon Source (APS). Access to the operation of these beamlines is accomplished through visits by investigators who are either on-site, remote or a combination of the two. In all modes of access, user operations are controlled by the experimenter. The control and capabilities of the GM/CA beamlines are identical for remote and on-site users. Remote access to the beamlines is through NX or Teamviewer to local computers [1]. Once communication has been established, experienced GM/CA experimenters are greeted by our familiar JBluIce, the graphical user interface/control program[2] responsible for all operations from sample handling through data collection and reduction. Although investigators always see a familiar interface, both software and hardware on the beamlines are continually improving. Recent hardware upgrades include a shift of the optical focusing mirrors on the ID-B beamline closer to the sample to provide a significant increase in flux, and installation of a new Pilatus3 6M detector on ID-D, the second undulator beamline. The JBluIce program has incorporated new detector controls for shutterless operation while continuing to expand the features of rastering, vector (helical) data collection, strategy tools and data analysis. These tools have been essential to investigators working on membrane crystal samples, e.g. GPCRs, as well as for samples that decay quickly or require data to be collected from multiple crystals. The presentation will provide an overview of beamline remote control as well as an update of the equipment that it operates at GM/CA.