for advanced facility: SAXS Studies on Structures of Biological - PowerPoint PPT Presentation

Current status of 4C SAXS beamline and its long-term plan for advanced facility: SAXS Studies on Structures of Biological Macromolecules in Solution at 4C Beamline Kyeong Sik Jin* & Byoungseok Min Pohang Accelerator Laboratory, Pohang University of Science & Technology, Pohang 790-784, Republic of Korea Email: jinks@postech.ac.kr Polymer Synthesis & Physics Laboratory

Contents 1. Introduction of 4C SAXS Beamline at PAL 2. SAXS Data Treatment and Analysis Procedures [Skip] 3. Research Results of Biomacromolecules in Solution

2016 PLS – II Operation Schedule and Status

Beamline Map and Status 30 operation BLs, 10 ID BLs, 3 ID + 1 Bending SAXS BLs (Material Chemistry)

4C SAXS BL : Small-angle X-ray Scattering Small-angle X-ray Scattering (SAXS) is a small-angle scattering (SAS) technique where the elastic scattering of X-rays (wavelength 0.1 ~ 0.2 nm) by a sample which has inhomogeneities in the nm-range, is recorded at very low angle (typically 0.1 ~ 10 ° ). This angular range contains information about the shape and size of macromolecules, characteristic distances of partially ordered materials, pore sizes, and other data. SAXS is capable of delivering structural information of macromolecules between 3 and 25 nm, of repeat distances in partially ordered systems of up to 150 nm. U-SAXS (ultra-small angle X- ray scattering) can resolves even larger dimensions. 4C SAXS beamline is dedicated to conventional transmission small-angle X-ray scattering and diffraction studies for interpreting the structure, structural changes, and relationship between structure and function of molecular/nano structured polymer, self-assembled organic/inorganic nanostructure, composite nanomaterials, biological macromolecules (protein, DNA, and RNA) and their complexes in nearly physiological environments.

Applications The method is accurate, non-destructive and usually requires only a minimum of sample preparation. Applications are very broad and include colloids of all types, metals, cement, oil, polymers, plastics, proteins, foods and pharmaceuticals can be found in research as well as in quality control. SAXS is used for the determination of nanoscale structure of particle systems • Synthetic Polymer Nanostructure Studies: Drug Delivery System, Electronic Device • Biological Nanostructure Studies: Disease, Drug, Molecular Device • Energy Material: Organic Solar Cell, Battery • Flexible Display: Organic Thin-film Transistor • Ultra Low-k Dielectric Material • Nano-template Fabrication • Micro-array Fabrication SAXS is the universal method for nanostructure analysis • Sensitive – low concentration (down to 0.1 %) • Noninvasive – no preparation, staining or drying • In -situ, real-time capabilities • Extremely versatile (solids, liquids, gas phase)

Resolution

Scattering Intensity Form factor F(q)   I ( q ) F ( q ) S ( q ) S(q)  1 For BioSAXS, size, shape, orientation log I (a.u.) I(q) S(q) Structure factor S(q) F(q) 0.0 0.5 1.0 1.5 local order, relative position

X-ray Scattering: SAXS vs. WAXS? SAXS patterns contain data concerning correlations on an inter-molecular level: necessary samples where there is macromolecular or aggregate order WAXS patterns contain data concerning correlations on an intra-molecular, inter- atomic level As synthesis design/control improves, SAXS becomes more relevant than ever before

4C SAXS II Beamline Layout Incident beam height = 1400 mm Experimental Hutch Optical Hutch Front End Movable Beam Undulator Wall mask stopper 88.4 mm Incidence Angle 2.6 mrad (including graphite filter) (including copper screen/tungsten wire monitors) Horizontal (33.00 urad) Beam Size [mm] 1.030 1.450 1.650 300 um Vertical (29.00 urad) Beam Size [mm] 0.400 0.700 0.700 23 um Source Defining Slit DCM FM Detector 0 m 11.4 m 18.0 m 21.3 m 37 m

Experimental Components in Hutch

A Brief Introduction about 4C SAXS II Beamline Beamline Specifications Previous and Current Status Development and construction of EPICS-based software  Beamline 4C SAXS-II package for electronic optical component and Status Operational experimental equipment control (Self-development) Development and construction of user friendly  In-vacuum Undulator 20 Source integrated software package for data measurement and (1.4 m short, 20 mm period) data treatment (Self-development) Monochromator DCM Si (111) Design and fabrication of a variety of sample cell &  stage systems for bulk/powder/film/solution SAXS and 0.06-0.12 nm, Wavelength WAXD experiments (Self-development) currently 0.07 nm Construction of experimental equipments for sample  Vertical focusing toroidal, Mirror storage, preparation, treatment, and basic material rhodium coated property analysis 1 × 10 12 ph/sec Beam flux Activation of user groups and of joint researches  100(V) × 300(H) µm 2 Beam size Resolution 200 nm ~ 0.3 nm Future Projects and Goals Individually motorized blades of Slits tungsten (W) Sample-to-detector  Further development of methods and approaches for 5.0/4.0/3.0/2.0/1.0/0.5/0.2 m distance high-throughput application measurements, which is available at 4C beamline Detector Rayonix 2D SX 165  Further improvement of the performance of 4C Bulk, solution, liquid crystal, film, beamline, including automation of SAXS experiments Experimental methods powder, sol-gel T-SAXS, T-WAXD and data analysis, and time-resolved SAXS setups  Embarkation on a collaborative research project to 50 user groups (Polymer, bio-, User groups study the structure of a wide range of self-assembled organic, inorganic groups) nanostructures and biological macromolecules Research results 40 ~ 50 papers/year  Construction of computer system for molecular dynamic (MD) simulation that could be combined well with time- ATSAS package, SCATTER, etc. Data analysis software resolved SAXS study – Pre-dynamic SAXS studies aimed (free download) at the 4 th generation XFEL experiment SAXS studies for self-assembled Staff science nanostructures and biomacromolecules in solution

4C Small angle X-ray scattering (SAXS) Beamline at PAL Sample Environments: ● Multi (100 holes) Sample Stage for Powder, Bulk, Film (25 ºC) ● Multi (10 lines) Sample Stage for Scan of Film (25 ºC) ● Multi (6 holes) Heating Sample Stage equipped with Eurotherm Controller for Powder, Bulk, Film (25 ~ 400 ºC) ● Multi (5 holes) Cooling/Heating Sample Stage equipped with Julabo Circulation for Solution (-15 ~ 200 ºC) ● Multi (5 holes) Heating Sample Stage equipped with Eurotherm Controller for Solution (25 ~ 400 ºC) ● Single Cooling/Heating Sample Stage for Sol-gel (-15 ~ 400 ºC) Equipments: 1)Malvern Zetasizer Nano Series DLS, 2)Refrigerated/multipurpose/high speed Centrifuge, 3)Ultarsonic Cleaner, 4)Water Purification System, 5)Cold Storage (4ºC), 6)Ultra-Low Temperature Freezer (-40ºC), 7)Bench Mixer, 8)Hanil Micro-12, 9)Thermo Scientific Heater, 10)METTLER TOLEDO EL204-IC Electronic Balance, 11)Pipettes, 12)S-1700, 13)NanoDrop, 14)GPC-FFF-MALS System

Sample Environments Multi (10 lines) sample stage for film (25 ºC) Multi (100 holes) sample stage for powder, bulk, film (25 ºC) Temperature sensor Sample cell N2 inlet Heating bar cable Cooling water inlet Multi (6 holes) heating sample stage equipped with Single heating sample stage equipped with Eurotherm Eurotherm Controller for powder, bulk, film (25 ~ 400 ºC) Controller for powder, bulk, film (25 ~ 400 ºC)

Sample Environments Sample cell Julabo Pipette + Tip Quartz capillary connection part heating bar cable Single cooling/heating sample stage for sol-gel (-15 ~ 400 ºC) Quartz-based solution cell Single cooling/heating sample stage equipped with Julabo circulation Multi (20 holes) cooling/heating sample stage for sol-gel (-15 ~ 200 ºC) for solution (-15 ~ 200 ºC)

Dual Detector System for Rayonix 165 and Pilatus 1M

GPC-FFF-MALS System World-class Beamline UV/Vis Pump MALS GPC (18 channel) column RI Sample FFF injector channel Fraction FFF collector [FFF] 1) Injection flow After Before 3) Channel flow 2) Cross flow

GPC-FFF-MALS System ◈ Test Result of BSA Protein Standard (MW = 66.450kDa, C= 2mg/ml, Injection volume = 15 ㎕ , Experiment time = 13 min)

EPICS IOC-based Control for Main Optics STEPⅠ

Python-based Data Treatment Software Automatic/selective background - transmission corrected 2D data acquisition - 1D radial averaged data acquisition – save, batch processing program buildup (1) (4) (6) (3) (2) (5) STEP Ⅱ

A World-wide Leading Beamline ◈ Bio SAXS P12 beamline at the Petra- Ⅲ storage ring (Leader: Dr. Dmitri Svergun) Purification system Dynamic and movable detector Automatic sample changer