Small Angle Scattering (SAXS/SANS) Small Angle Scattering - PowerPoint PPT Presentation

European Molecular Biology Laboratory Hamburg Outstation European Molecular Biology Laboratory Hamburg Outstation European Molecular Biology Laboratory Hamburg Outstation European Molecular Biology Laboratory Hamburg Outstation BioSAXS group BioSAXS group BioSAXS group BioSAXS group Small Angle Scattering (SAXS/SANS) Small Angle Scattering (SAXS/SANS) Small Angle Scattering (SAXS/SANS) Small Angle Scattering (SAXS/SANS) data reduction and overall parameters data reduction and overall parameters data reduction and overall parameters data reduction and overall parameters Dr. Weifeng Shang 尚伟峰 Dr. Weifeng Shang Dr. Weifeng Shang Dr. Weifeng Shang

Outline Outline Outline Outline 1 Sample preparation 1 Sample preparation 1 Sample preparation 1 Sample preparation 2 SAXS in theory 2 SAXS in theory 2 SAXS in theory 2 SAXS in theory 3 SAXS in practice 3 SAXS in practice 3 SAXS in practice 3 SAXS in practice 4 Data collection 4 Data collection 4 Data collection 4 Data collection 5 Data reduction 5 Data reduction 5 Data reduction 5 Data reduction 6 Overall parameters 6 Overall parameters 6 Overall parameters 6 Overall parameters

Sample preparation Sample preparation Sample preparation Sample preparation Exact sequence sequence sequence sequence ---> predicted molecular mass molecular mass molecular mass molecular mass True concentration True concentration True concentration True concentration ( extinction coefficient, absorbance at 280 nm ) Mono-disperse samples Mono-disperse samples Mono-disperse samples Mono-disperse samples ( precipitate, SDS-PAGE, GF/SE chromatography, DLS, AUC, mass spectrometer )

Buffer preparation Buffer preparation Buffer preparation Buffer preparation Matched buffer Matched buffer Matched buffer Matched buffer ( the exact buffer at dialysis equilibrium with the sample ) ( buffer exchange, dialysis ) Good buffer Good buffer Good buffer Good buffer 1. Tris, Hepes, PBS, HBS, etc. 2. salts, additives, small molecules, DTT 3. detergents, < CMC

SAXS in theory SAXS in theory SAXS in theory SAXS in theory

SAXS in practice SAXS in practice SAXS in practice SAXS in practice

Data collection Data collection Data collection Data collection Commonly used detectors for SAXS (0.15 nm, 8.3 keV) Commonly used detectors for SAXS (0.15 nm, 8.3 keV) Commonly used detectors for SAXS (0.15 nm, 8.3 keV) Commonly used detectors for SAXS (0.15 nm, 8.3 keV) 1. Position sensitive gas-filled proportional counter 1. Position sensitive gas-filled proportional counter 1. Position sensitive gas-filled proportional counter 1. Position sensitive gas-filled proportional counter 2. PILATUS detector 2. PILATUS detector 2. PILATUS detector 2. PILATUS detector 3. Image plate 3. Image plate 3. Image plate 3. Image plate 4. CCD detector 4. CCD detector 4. CCD detector 4. CCD detector

Data reduction Data reduction Data reduction Data reduction

Data reduction Data reduction Data reduction Data reduction 1 (buffer, sample, buffer) sequence 1 1 (buffer, sample, buffer) sequence 1 (buffer, sample, buffer) sequence (buffer, sample, buffer) sequence 2 2 2 Radial integration (2D 1D) 2 Radial integration (2D 1D) Radial integration (2D 1D) Radial integration (2D 1D) 3 3 Check for radiation damage Check for radiation damage 3 3 Check for radiation damage Check for radiation damage 4 4 Average frames Average frames 4 4 Average frames Average frames 5 Buffer averaging 5 Buffer averaging 5 Buffer averaging 5 Buffer averaging 6 Buffer subtraction 6 Buffer subtraction 6 Buffer subtraction 6 Buffer subtraction

Data reduction Data reduction Data reduction Data reduction

Data reduction Data reduction Data reduction Data reduction: buffer averaging : buffer averaging : buffer averaging : buffer averaging

Data reduction Data reduction Data reduction Data reduction: buffer subtraction : buffer subtraction : buffer subtraction : buffer subtraction

Overall parameters: Rg Overall parameters: Rg Overall parameters: Rg Overall parameters: Rg

Overall parameters: AutoRg Overall parameters: AutoRg Overall parameters: AutoRg Overall parameters: AutoRg

Overall parameters: Porod Volume Overall parameters: Porod Volume Overall parameters: Porod Volume Overall parameters: Porod Volume

Overall parameters: GNOM Overall parameters: GNOM Overall parameters: GNOM Overall parameters: GNOM

Overall parameters: p(r) Overall parameters: p(r) Overall parameters: p(r) Overall parameters: p(r)

Overall parameters: Dmax Overall parameters: Dmax Overall parameters: Dmax Overall parameters: Dmax

SAXS pipeline SAXS pipeline SAXS pipeline SAXS pipeline

Small Angle Scattering (SAXS/SANS) Small Angle Scattering - PowerPoint PPT Presentation

European Molecular Biology Laboratory Hamburg Outstation European Molecular Biology Laboratory Hamburg Outstation European Molecular Biology Laboratory Hamburg Outstation European Molecular Biology Laboratory Hamburg Outstation BioSAXS group

Joint Application of SAXS and SANS Jill Trewhella University of Sydney Small-angle scattering of

Overview L8.1 Introduction to Small Angle Neutron Scattering L8.2 SANS Instrumentation

Overview L8.1 Introduction to Small Angle Neutron Scattering L8.2 SANS Instrumentation

Sample Preparation and Characterization Cy Jeffries EMBL Hamburg Small-angle scattering (SAS)

SAXS and SANS facilities and experimental practice Clement Blanchet EMBL Hamburg Small Angle

SAXS and SANS facilities and experimental practice Clement Blanchet EMBL Hamburg Small Angle

Ab initio methods: how/why do they work D.Svergun, EMBL-Hamburg Small- -angle scattering in

Scattering of Neutrons: Basics Jill Trewhella University of Sydney Small-angle scattering of

Ab initio methods: how/why do they work D.Svergun Small-angle scattering in structural biology

Small angle X-ray scattering from biological macromolecules in solution Al Kikhney European

SAXS/ SANS data processing and overall parameters Petr V. Konarev European Molecular Biology

X-ray reflectivity and Grazing Incidence Small Angle X-Ray Scattering G. Renaud, CEA-Grenoble,

Complementary use of SAXS and SANS Complementary use of SAXS and SANS Jill Trewhella Trewhella

Ab initio methods: how/why do they work Ab i iti th d h / h d th k D.Svergun Small Small-

Determination of apoferritin structure parameters using small angle neutron scattering Daniela

Combined use of small-angle X-ray scattering and functional studies: the case of glutamate

Particle Growth in Mixed Matrix Polymer Membranes: Using Ultra-Small Angle Neutron Scattering to

SASBDB Small Angle Scattering Biological Data Bank Erica Valentini Dmitri Svergun group

Small Angle Scattering Ab initio modelling Requirements for Data Collection sample must be

small angle scattering data Al Kikhney EMBL Hamburg, Germany Outline Validation of atomic

Status Micro-focused Small Angle Neutron Scattering CRP At RTP Presented at Presented at

Scattering - Introduction We will consider two types of scattering Scattering on

Geant4 implementation of inter-atomic interference effect in Small-Angle Coherent X-ray

WAXS going beyond SAXS Lee Makowski Northeastern University Boston WAXS (wide angle