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. 2022:677:157-189.
doi: 10.1016/bs.mie.2022.08.008. Epub 2022 Sep 22.

Technical considerations for small-angle neutron scattering from biological macromolecules in solution: Cross sections, contrasts, instrument setup and measurement

Zuzanna Pietras  1 , Kathleen Wood  2 , Andrew E Whitten  3 , Cy M Jeffries  4
Affiliations

Technical considerations for small-angle neutron scattering from biological macromolecules in solution: Cross sections, contrasts, instrument setup and measurement

Zuzanna Pietras et al. Methods Enzymol. 2022.

Abstract

Small angle scattering affords an approach to evaluate the structure of dilute populations of macromolecules in solution where the measured scattering intensities relate to the distribution of scattering-pair distances within each macromolecule. When small angle neutron scattering (SANS) with contrast variation is employed, additional structural information can be obtained regarding the internal organization of biomacromolecule complexes and assemblies. The technique allows for the components of assemblies to be selectively 'matched in' and 'matched out' of the scattering profiles due to the different ways the isotopes of hydrogen-protium 1H, and deuterium 2H (or D)-scatter neutrons. The isotopic substitution of 1H for D in the sample enables the controlled variation of the scattering contrasts. A contrast variation experiment requires trade-offs between neutron beam intensity, q-range, wavelength and q-resolution, isotopic labelling levels, sample concentration and path-length, and measurement times. Navigating these competing aspects to find an optimal combination is a daunting task. Here we provide an overview of how to calculate the neutron scattering contrasts of dilute biological macromolecule samples prior to an experiment and how this then informs the approach to configuring SANS instruments and the measurement of a contrast variation series dataset.

Keywords: Biological macromolecule; Contrast; Contrast variation; Dilute solution scattering; SANS; SANS instrument.

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