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Title: Solvent minimization induces preferential orientation and crystal clustering in serial micro-crystallography on micro-meshes, in situ plates and on a movable crystal conveyor belt

Journal Article · · Journal of Synchrotron Radiation (Online)
 [1];  [2];  [3];  [1];  [1];  [1];  [1];  [1];  [1];  [1]
  1. Brookhaven National Lab. (BNL), Upton, NY (United States)
  2. Brookhaven National Lab. (BNL), Upton, NY (United States); Univ. of Oregon, Eugene, OR (United States) Dept. of Physics
  3. Brookhaven National Lab. (BNL), Upton, NY (United States); Suffolk County Community College, Selden, NY (United States)
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X-ray diffraction data were obtained at the National Synchrotron Light Source from insulin and lysozyme crystals that were densely deposited on three types of surfaces suitable for serial micro-crystallography: MiTeGen MicroMeshes™, Greiner Bio-One Ltdin situmicro-plates, and a moving kapton crystal conveyor belt that is used to deliver crystals directly into the X-ray beam. 6° wedges of data were taken from ~100 crystals mounted on each material, and these individual data sets were merged to form nine complete data sets (six from insulin crystals and three from lysozyme crystals). Insulin crystals have a parallelepiped habit with an extended flat face that preferentially aligned with the mounting surfaces, impacting the data collection strategy and the design of the serial crystallography apparatus. Lysozyme crystals had a cuboidal habit and showed no preferential orientation. Preferential orientation occluded regions of reciprocal space when the X-ray beam was incident normal to the data-collection medium surface, requiring a second pass of data collection with the apparatus inclined away from the orthogonal. In addition, crystals measuring less than 20 µm were observed to clump together into clusters of crystals. Clustering required that the X-ray beam be adjusted to match the crystal size to prevent overlapping diffraction patterns. No additional problems were encountered with the serial crystallography strategy of combining small randomly oriented wedges of data from a large number of specimens. Lastly, high-quality data able to support a realistic molecular replacement solution were readily obtained from both crystal types using all three serial crystallography strategies.

Research Organization:
Brookhaven National Laboratory (BNL), Upton, NY (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
Grant/Contract Number:
SC00112704; 11- 008; P41RR012408; P41GM103473
OSTI ID:
1229355
Report Number(s):
BNL-111430-2015-JA; JSYRES
Journal Information:
Journal of Synchrotron Radiation (Online), Vol. 21, Issue 6; ISSN 1600-5775
Publisher:
International Union of CrystallographyCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 10 works
Citation information provided by
Web of Science

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Cited By (3)

Microfluidics: From crystallization to serial time-resolved crystallography journal May 2017
On-chip crystallization for serial crystallography experiments and on-chip ligand-binding studies text January 2019
On-chip crystallization for serial crystallography experiments and on-chip ligand-binding studies text January 2019

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Related Subjects

36 MATERIALS SCIENCE
in situ X-ray data collection
crystallography
acoustic droplet ejection
serial crystallography