Efficient in situ screening of and data collection from microcrystals in crystallization plates.

IF 2.6 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS
Amy J Thompson, Juan Sanchez-Weatherby, Lewis J Williams, Halina Mikolajek, James Sandy, Jonathan A R Worrall, Michael A Hough
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引用次数: 0

Abstract

A considerable bottleneck in serial crystallography at XFEL and synchrotron sources is the efficient production of large quantities of homogenous, well diffracting microcrystals. Efficient high-throughput screening of batch-grown microcrystals and the determination of ground-state structures from different conditions is thus of considerable value in the early stages of a project. Here, a highly sample-efficient methodology to measure serial crystallography data from microcrystals by raster scanning within standard in situ 96-well crystallization plates is described. Structures were determined from very small quantities of microcrystal suspension and the results were compared with those from other sample-delivery methods. The analysis of a two-dimensional batch crystallization screen using this method is also described as a useful guide for further optimization and the selection of appropriate conditions for scaling up microcrystallization.

Abstract Image

对结晶板中的微晶体进行高效的原位筛选和数据采集。
在 XFEL 和同步辐射源上进行系列晶体学研究的一个相当大的瓶颈是如何有效地生产大量同质、衍射良好的微晶体。因此,对批量生产的微晶体进行高效的高通量筛选,并确定不同条件下的基态结构,在项目的早期阶段具有相当大的价值。本文介绍了一种在标准原位 96 孔结晶板内通过光栅扫描测量微晶体序列结晶数据的高样品效率方法。从极少量的微晶悬浮液中确定了晶体结构,并将结果与其他样品输送方法进行了比较。还介绍了使用这种方法对二维批量结晶筛选进行的分析,为进一步优化和选择扩大微晶规模的适当条件提供了有用的指导。
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来源期刊
Acta Crystallographica. Section D, Structural Biology
Acta Crystallographica. Section D, Structural Biology BIOCHEMICAL RESEARCH METHODSBIOCHEMISTRY &-BIOCHEMISTRY & MOLECULAR BIOLOGY
CiteScore
4.50
自引率
13.60%
发文量
216
期刊介绍: Acta Crystallographica Section D welcomes the submission of articles covering any aspect of structural biology, with a particular emphasis on the structures of biological macromolecules or the methods used to determine them. Reports on new structures of biological importance may address the smallest macromolecules to the largest complex molecular machines. These structures may have been determined using any structural biology technique including crystallography, NMR, cryoEM and/or other techniques. The key criterion is that such articles must present significant new insights into biological, chemical or medical sciences. The inclusion of complementary data that support the conclusions drawn from the structural studies (such as binding studies, mass spectrometry, enzyme assays, or analysis of mutants or other modified forms of biological macromolecule) is encouraged. Methods articles may include new approaches to any aspect of biological structure determination or structure analysis but will only be accepted where they focus on new methods that are demonstrated to be of general applicability and importance to structural biology. Articles describing particularly difficult problems in structural biology are also welcomed, if the analysis would provide useful insights to others facing similar problems.
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