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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.

ABSTRACT: 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 Ltd in situ micro-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. High-quality data able to support a realistic molecular replacement solution were readily obtained from both crystal types using all three serial crystallography strategies.

SUBMITTER: Soares AS

PROVIDER: S-EPMC4211130 | biostudies-literature | 2014 Nov

REPOSITORIES: biostudies-literature

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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.

Soares Alexei S AS Mullen Jeffrey D JD Parekh Ruchi M RM McCarthy Grace S GS Roessler Christian G CG Jackimowicz Rick R Skinner John M JM Orville Allen M AM Allaire Marc M Sweet Robert M RM

Journal of synchrotron radiation 20141009 Pt 6

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 Ltd in situ micro-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 ...[more]

PMID: 25343789

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Project description:During automated processing in commercial hatcheries, day-old chicks are subjected to a range of possible mental and physical stressors. Three determinants of the processing line seem to have the potential to affect the birds in particular: drop height from one conveyor belt to another, conveyor belt speed, and acceleration. The aim of this study was to evaluate the effects of these 3 factors on chicken health and welfare in early and later life. In a first trial, chickens were tested on an experimental processing line that was adjusted to different levels of drop heights, belt speeds, and accelerations separately (n = 14 animals per factor and increment). Besides the assessment of several indicators for disorientation during the treatment, postmortem radiographic images were created and analyzed with focus on traumatic injuries. The number of chickens changing their orientation after the drop was affected by drop height (P < 0.01), whereas body posture changes were affected both by drop height (P < 0.01) and belt speed (P < 0.01). Traumatic injuries were found only sporadically and were not related to a certain treatment. In a second trial, chickens that were exposed to a combination of the 3 processing factors were compared with an untreated control group (n = 63 per group) until 15 d of age. There were no differences between the 2 groups regarding BW, welfare scores, and fear-related responses in a novel object and in a tonic immobility test. The present results suggest that the treatments on the experimental conveyor belts affected the birds' health, welfare, and behavior to a limited extend. However, starting at a drop height of 280 mm and a conveyor belt speed of 27 m/min, significantly more chickens were not able to maintain their initial body position on the belt. This indicates that there may be scope for discomfort and welfare impairment if commercial systems are operated with considerably larger drop heights and at higher speeds.

Dataset Information

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.

Publications

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.

Similar Datasets

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