Depth-of-field extension in optical imaging for rapid crystal screening
Depth-of-field extension in optical imaging for rapid crystal screening
复制标题
用于快速晶体筛选的光学成像景深扩展
DOI：
10.1107/s2059798321000097
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Simpson, Garth
中科院分区：
文献类型：
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作者：
Li, Chen;Ding, Changqin;Li, Minghe;Rong, Jiayue;Florian, Hilary;Simpson, Garth
通过在非线性光学显微镜的光路中改装定制设计的微延迟器阵列（µRA），景深（DoF）扩展了2.8倍，以实现快速晶体筛选。在蛋白质晶体的二次谐波生成成像的应用中，评估了所提出的DoF增强策略的优点。结果发现，DoF延伸增加了检测到的晶体数量，同时减少了筛选所需的“z切片”数量。实验测量的扩展自由度的波长依赖性与理论预测非常吻合。这些结果提供了一个简单和广泛适用的方法，以提高现有的非线性光学成像方法的蛋白质晶体筛选的吞吐量。
The depth of field (DoF) was extended 2.8-fold to achieve rapid crystal screening by retrofitting a custom-designed micro-retarder array (µRA) in the optical beam path of a nonlinear optical microscope. The merits of the proposed strategy for DoF enhancement were assessed in applications of second-harmonic generation imaging of protein crystals. It was found that DoF extension increased the number of crystals detected while simultaneously reducing the number of `z-slices' required for screening. Experimental measurements of the wavelength-dependence of the extended DoF were in excellent agreement with theoretical predictions. These results provide a simple and broadly applicable approach to increase the throughput of existing nonlinear optical imaging methods for protein crystal screening.