SLIDE 1 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
- Dr. Weifeng Shang
- Dr. Weifeng Shang
- Dr. Weifeng Shang 尚伟峰
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
SLIDE 2
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
SLIDE 3
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)
SLIDE 4 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
SLIDE 5
SAXS in theory SAXS in theory SAXS in theory SAXS in theory
SLIDE 6
SAXS in practice SAXS in practice SAXS in practice SAXS in practice
SLIDE 7 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
SLIDE 8
Data reduction Data reduction Data reduction Data reduction
SLIDE 9
Data reduction Data reduction Data reduction Data reduction
SLIDE 10
Data reduction Data reduction Data reduction Data reduction
SLIDE 11
Data reduction Data reduction Data reduction Data reduction
SLIDE 12
1 1 1 1 (buffer, sample, buffer) sequence (buffer, sample, buffer) sequence (buffer, sample, buffer) sequence (buffer, sample, buffer) sequence 2 2 2 2 Radial integration (2D 1D) Radial integration (2D 1D) Radial integration (2D 1D) Radial integration (2D 1D) 3 3 3 3 Check for radiation damage Check for radiation damage Check for radiation damage Check for radiation damage 4 4 4 4 Average frames Average frames 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
SLIDE 13
Data reduction Data reduction Data reduction Data reduction
SLIDE 14
Data reduction Data reduction Data reduction Data reduction: buffer averaging : buffer averaging : buffer averaging : buffer averaging
SLIDE 15
Data reduction Data reduction Data reduction Data reduction: buffer averaging : buffer averaging : buffer averaging : buffer averaging
SLIDE 16
Data reduction Data reduction Data reduction Data reduction: buffer subtraction : buffer subtraction : buffer subtraction : buffer subtraction
SLIDE 17
Data reduction Data reduction Data reduction Data reduction: buffer subtraction : buffer subtraction : buffer subtraction : buffer subtraction
SLIDE 18
Overall parameters: Rg Overall parameters: Rg Overall parameters: Rg Overall parameters: Rg
SLIDE 19
Overall parameters: AutoRg Overall parameters: AutoRg Overall parameters: AutoRg Overall parameters: AutoRg
SLIDE 20
Overall parameters: Porod Volume Overall parameters: Porod Volume Overall parameters: Porod Volume Overall parameters: Porod Volume
SLIDE 21
Overall parameters: GNOM Overall parameters: GNOM Overall parameters: GNOM Overall parameters: GNOM
SLIDE 22
Overall parameters: p(r) Overall parameters: p(r) Overall parameters: p(r) Overall parameters: p(r)
SLIDE 23
Overall parameters: p(r) Overall parameters: p(r) Overall parameters: p(r) Overall parameters: p(r)
SLIDE 24
Overall parameters: p(r) Overall parameters: p(r) Overall parameters: p(r) Overall parameters: p(r)
SLIDE 25
Overall parameters: Dmax Overall parameters: Dmax Overall parameters: Dmax Overall parameters: Dmax
SLIDE 26
Overall parameters: Dmax Overall parameters: Dmax Overall parameters: Dmax Overall parameters: Dmax
SLIDE 27
Overall parameters: Dmax Overall parameters: Dmax Overall parameters: Dmax Overall parameters: Dmax
SLIDE 28
SAXS pipeline SAXS pipeline SAXS pipeline SAXS pipeline
