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$Measurements of Long-range Electronic Correlations During Femtosecond Diffraction Experiments Performed on Nanocrystals of Buckminsterfullerene$

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Considerations for three-dimensional image reconstruction from experimental data in coherent diffractive imaging.

Ida V Lundholm¹, Jonas A Sellberg², Tomas Ekeberg¹

¹Laboratory of Molecular Biophysics, Department of Cell and Molecular Biology, Uppsala University, Husargatan 3 (Box 596), SE-751 24 Uppsala, Sweden.

|September 19, 2018

Summary

This summary is machine-generated.

Diffraction before destruction with X-ray free-electron lasers (XFELs) enables radiation-damage-free imaging. Background noise in diffraction patterns creates central artifacts, but background removal and advanced algorithms can improve structural reconstructions.

Keywords:

LCLS Melbournevirus XFELs coherent diffractive imaging image reconstruction

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Area of Science:

Structural biology
Biophysics
X-ray crystallography

Background:

Diffraction before destruction (DBD) using X-ray free-electron lasers (XFELs) offers a method for radiation-damage-free structural determination.
Crystallization is not required for this technique, making it suitable for challenging samples.

Purpose of the Study:

To perform three-dimensional reconstruction of the Melbourne virus using single-particle X-ray diffraction.
To investigate the impact of experimental noise on structural reconstructions from XFEL data.

Main Methods:

Collected single-particle X-ray diffraction patterns from Melbourne virus at the Linac Coherent Light Source (LCLS).
Generated simulated diffraction data to explore the effects of various noise sources, including experimental background and blurring.
Applied phase-retrieval algorithms and Fourier-volume-assembly techniques.

Main Results:

A significant central artifact was observed in the experimental reconstruction, which was reproducible in simulations with added background noise.
The artifact's intensity correlated linearly with background strength, indicating its origin from the Fourier transform of the background.
Artifacts were reduced by implementing background removal prior to phase retrieval.
Blurring introduced diffuse artifacts mistaken for biological features, while sample heterogeneity and pulse energy variations had minimal impact.

Conclusions:

Experimental background is a primary source of artifacts in XFEL single-particle imaging, particularly central features.
Background removal and advanced reconstruction algorithms are crucial for accurate structural determination.
High repetition-rate XFELs and background-aware assembly algorithms will enhance signal-to-noise ratios and minimize artifacts in future studies.