eCommons

 

Exploration of Methods for Serial Microcrystallography at Storage Ring X-ray Sources

Other Titles

Abstract

Protein crystallography has made the largest contribution to our knowledge of protein structure. However, it is well known that many biologically important proteins do not readily form large enough crystals for traditional crystallography. Successful serial microcrystallography (SMX) studies have been performed at X-ray Free-Electron Lasers, but they are limited in experimental availability. We look to more accessible light sources, such as storage ring sources, for the development of SMX. However, improvements to the conventional experiment are required to make SMX viable at storage rings. Here, we explore several devices and techniques designed given this consideration. To isolate crystal diffraction, the sample environment should contribute zero background scatter outside the crystal, since excess scatter obscures the weak microcrystal signal. We will show the feasibility of using atomically-thin, gas-tight graphene to reduce background scatter as a crystal mount and suggest using it as a window material for SMX. We will also explore two microcrystal delivery devices, a microfluidic chip and a viscous jet injector, for use in SMX. While both of these devices show promise for optimizing various aspects of the crystal delivery system, both contribute more background scatter than is acceptable for an ideal SMX experiment at a storage ring source. Merging diffraction from multiple microcrystals is necessary when a complete data set cannot be determined from a single microcrystal. When microcrystal diffraction is weak enough that Bragg peaks are no longer visible, merging through conventional techniques fails since crystal orientation cannot be obtained through Bragg peak indexing. We will explore proof-of-principle experiments which show that indexing data frames on a per-frame basis is unnecessary for a structure solution, when reciprocal space intensities can be reconstructed using the EMC algorithm. In principle, serial microcrystallography is feasible at storage ring sources if improvements in beamline setups, sample chamber construction and microcrystal diffraction analysis evolve to optimize the diffraction of microcrystals.

Journal / Series

Volume & Issue

Description

Sponsorship

Date Issued

2017-01-30

Publisher

Keywords

microcrystallography; serial crystallography; sparse data; synchrotron storage ring sources; x-ray protein crystallography; Biophysics; EMC algorithm

Location

Effective Date

Expiration Date

Sector

Employer

Union

Union Local

NAICS

Number of Workers

Committee Chair

Gruner, Sol Michael

Committee Co-Chair

Committee Member

Crane, Brian
Pollack, Lois

Degree Discipline

Biophysics

Degree Name

Ph. D., Biophysics

Degree Level

Doctor of Philosophy

Related Version

Related DOI

Related To

Related Part

Based on Related Item

Has Other Format(s)

Part of Related Item

Related To

Related Publication(s)

Link(s) to Related Publication(s)

References

Link(s) to Reference(s)

Previously Published As

Government Document

ISBN

ISMN

ISSN

Other Identifiers

Rights

Attribution 4.0 International

Types

dissertation or thesis

Accessibility Feature

Accessibility Hazard

Accessibility Summary

Link(s) to Catalog Record