Development of high-performance X-ray transparent crystallization plates for in situ protein crystal screening and analysis.
Development of high-performance X-ray transparent crystallization plates for in situ protein crystal screening and analysis.
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DOI：
10.1107/s090744491101883x
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发表时间：
2011-07
期刊：
影响因子：
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通讯作者：
A. Soliman;M. Warkentin;B. Apker;R. Thorne
中科院分区：
文献类型：
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作者：
A. Soliman;M. Warkentin;B. Apker;R. Thorne
基于新型滴钉技术的x射线透明结晶板为蛋白质结晶和筛选提供了一种灵活、简单和廉价的方法。该板由开放的细胞组成，顶部和底部由薄薄的光学，紫外线和x射线透明薄膜密封。平板不需要孔或凹陷来容纳液体。取而代之的是，蛋白质滴和储液通过微米尺寸的环固定在底部膜上。这些环牢固地固定了液体接触线，从而提高了液滴形状和位置的均匀性，从而提高了结晶重现性，简化了液滴含量的自动图像分析。同样的环有效地固定了含有盐、蛋白质、冷冻保护剂、油、醇和洗涤剂的溶液。强钉环允许板旋转没有液体混合到90°x射线数据收集或倒转挂滴结晶。该板具有标准的SBS格式，并与标准的液体处理机器人兼容。
X-ray transparent crystallization plates based upon a novel drop-pinning technology provide a flexible, simple and inexpensive approach to protein crystallization and screening. The plates consist of open cells sealed top and bottom by thin optically, UV and X-ray transparent films. The plates do not need wells or depressions to contain liquids. Instead, protein drops and reservoir solution are held in place by rings with micrometre dimensions that are patterned onto the bottom film. These rings strongly pin the liquid contact lines, thereby improving drop shape and position uniformity, and thus crystallization reproducibility, and simplifying automated image analysis of drop contents. The same rings effectively pin solutions containing salts, proteins, cryoprotectants, oils, alcohols and detergents. Strong pinning by rings allows the plates to be rotated without liquid mixing to 90° for X-ray data collection or to be inverted for hanging-drop crystallization. The plates have the standard SBS format and are compatible with standard liquid-handling robots.