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首页> 美国卫生研究院文献>Journal of Synchrotron Radiation >Integrated nonlinear optical imaging microscope for on-axis crystal detection and centering at a synchrotron beamline
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Integrated nonlinear optical imaging microscope for on-axis crystal detection and centering at a synchrotron beamline

机译：集成式非线性光学成像显微镜用于同轴晶体检测并以同步加速器光束线为中心

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Nonlinear optical (NLO) instrumentation has been integrated with synchrotron X-ray diffraction (XRD) for combined single-platform analysis, initially targeting applications for automated crystal centering. Second-harmonic-generation microscopy and two-photon-excited ultraviolet fluorescence microscopy were evaluated for crystal detection and assessed by X-ray raster scanning. Two optical designs were constructed and characterized; one positioned downstream of the sample and one integrated into the upstream optical path of the diffractometer. Both instruments enabled protein crystal identification with integration times between 80 and 150 µs per pixel, representing a ∼103–104-fold reduction in the per-pixel exposure time relative to X-ray raster scanning. Quantitative centering and analysis of phenylalanine hydroxylase from Chromobacterium violaceum cPAH, Trichinella spiralis deubiquitinating enzyme TsUCH37, human κ-opioid receptor complex kOR-T4L produced in lipidic cubic phase (LCP), intimin prepared in LCP, and α-cellulose samples were performed by collecting multiple NLO images. The crystalline samples were characterized by single-crystal diffraction patterns, while α-cellulose was characterized by fiber diffraction. Good agreement was observed between the sample positions identified by NLO and XRD raster measurements for all samples studied.
机译：非线性光学（NLO）仪器已与同步加速器X射线衍射（XRD）集成在一起，用于组合式单平台分析，最初瞄准的是自动晶体居中的应用。对二次谐波产生显微镜和双光子激发紫外荧光显微镜进行晶体检测评估，并通过X射线光栅扫描进行评估。构造并表征了两种光学设计；一台位于样品的下游，另一台集成在衍射仪的上游光路中。两种仪器都可以进行蛋白质晶体鉴定，积分时间为每像素80至150μs，相对于每像素曝光时间而言，其减少了约10 3 –10 4 倍进行X射线光栅扫描。通过收集紫色小球藻cPAH，旋毛虫脱泛素化酶TsUCH37，在脂质立方相（LCP）中产生的人κ阿片受体复合物kOR-T4L，在LCP中制备的内膜素和α-纤维素样品中的苯丙氨酸羟化酶进行定量居中和分析多个NLO图像。晶体样品通过单晶衍射图表征，而α-纤维素通过纤维衍射表征。对于所有研究的样品，通过NLO和XRD栅格测量确定的样品位置之间均观察到良好的一致性。

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