SEC-SAXS Overview
In many cases, the solution SAXS data used for further analysis must be “clean” data in the sense that the scattering signals were obtained from monomodal and monodisperse sample. Even small amounts of higher oligomers and/or contaminants can interfere with the data analysis and make interpretation of the resulting scattering curves difficult and potentially misleading. Even if the sample of interest was initially obtained in monomodal and monodisperse state, many of problematic samples tend to aggregate rapidly prior to the SAXS measurement. Today, SEC-SAXS is a powerful tool to overcome such a heterogeneity problem. The SSRL BioSAXS beamline BL4-2 continues to develop the SEC-SAXS environment over the decade. A variety of experimental options are available for strategic SEC-SAXS data collection to overcome the two major challenges in biological SEC-SAXS experiments: the overlapping peaks in the elution profile and the fouling of radiation-damaged materials on the walls of the sample cell.
Update
Beamline SEC-SAXS paper has been published.
Beamline SEC-SAXS paper has been published.
Matsui T, Rajkovic I, Mooers BHM, Liu P, Weiss TM.
Adaptable SEC-SAXS data collection for higher quality structure analysis in solution.
Protein Sci. 2024 Apr;33(4):e4946. doi: 10.1002/pro.4946. PMID: 38501481; PMCID: PMC10949327.
Adaptable SEC-SAXS data collection for higher quality structure analysis in solution.
Protein Sci. 2024 Apr;33(4):e4946. doi: 10.1002/pro.4946. PMID: 38501481; PMCID: PMC10949327.
More details about BL4-2 SEC-SAXS are listed on the left sub-menu.
Main Features
■ Thermo Fisher Scientific UltiMate 3000 UHPLC
■ Automatic SEC-SAXS data collection and analysis (currently up to 50 data collections)
■ Optimizable X-ray energy and sample-detector distance for sample size and property
■ 3 SEC-SAXS modes are available:
- High Resolution mode 1)
- High-throughput mode (tandem operation using two SEC columns)
- Co-flow mode
■ Small internal volume to minimize sample dilution before/after SEC column
■ Additional electric buffer selection ports (currently maximum 8 buffer)
■ Small analytical SEC column allows data collection at higher sample concentration.
■ Sample economy: 2-100ul per injection 2)
■ Temperature controllable automatic sample injector and storage
■ Strategic SEC-SAXS data collection against undesired radiation damages
- Shutter control option (controlled by UV signal and/or elution time)
- Selection of X-ray positions on capillary
■ Further update on Blu-Ice SECSAXS tab, a beamline control software
■ Real-time automatic SEC-SAXS analytical pipeline, SECPipe
■ Customizable and powerful cleaning of capillary after every data collection
■ Additional capillary cleaning after background data collection
■ Multiple washing solutions and thorough washing by dispenser
■ Pilatus3 X 1M detector for noise-free data collection
■ Easily switchable to regular static measurement using SAXS Autosampler (TH-mode)
■ Multi-Angle Light Scattering detector, microDAWN (optional)
■ Refractive Index detector (optional)
■ Column jackets for the temperature control of the columns during analysis
1) Alternative to the High-Throughput or Co-flow mode.
2) We encourage the user to bring more volume of sample and buffer (see Sample Requirement for more details).