Three-dimensional single-particle imaging using angular correlations from X-ray laser data.

Femtosecond X-ray pulses from X-ray free-electron laser sources make it feasible to conduct room-temperature solution scattering experiments far below molecular rotational diffusion timescales. Owing to the ultra-short duration of each snapshot in these fluctuation scattering experiments, the particles are effectively frozen in space during the X-ray exposure. In contrast to standard small-angle scattering experiments, the resulting scattering patterns are anisotropic. The intensity fluctuations observed in the diffraction images can be used to obtain structural information embedded in the average angular correlation of the Fourier transform of the scattering species, of which standard small-angle scattering data are a subset. The additional information contained in the data of these fluctuation scattering experiments can be used to determine the structure of macromolecules in solution without imposing symmetry or spatial restraints during model reconstruction, reducing ambiguities normally observed in solution scattering studies. In this communication, a method that utilizes fluctuation X-ray scattering data to determine low-resolution solution structures is presented. The method is validated with theoretical data calculated from several representative molecules and applied to the reconstruction of nanoparticles from experimental data collected at the Linac Coherent Light Source.