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首页> 外文会议>Nanobiophotonics and Biomedical Applications III; Progress in Biomedical Optics and Imaging; vol.7, no.18 >Phase-Contrast BioCD: High-Speed Immunoassays at Sub-Picogram Detection Levels
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Phase-Contrast BioCD: High-Speed Immunoassays at Sub-Picogram Detection Levels

机译：相差BioCD：亚皮克级检测水平的高速免疫测定

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摘要

We previously introduced the biological compact disk (BioCD) as a sensitive detection platform to detect patterned biomolecules immobilized on the surface of a spinning disk. Spinning-disk interferometry allows high speed detection (10 microseconds per spot) of optical path length changes down to sub-nanometer scales with high repeatability. The key to performing stable interferometry on a mechanically spinning disk is self-referencing: locking the phase of the signal and reference beams to quadrature (π/2 phase difference) independent of mechanical vibrations or relative motion. Two quadrature classes of BioCD have been reported previously: the micro-diffraction class (MD-Class) and the adaptive optical class (AO-Class) {Peng, 2004 #565; Varma, 2004 #440}. In this paper, we introduce a third class of BioCD, the Phase-Contrast-Class (PC-Class) BioCD. Protein is immobilized using photolithography on a disk in a 1024 spoke pattern. The edge of the printed protein pattern diffracts a focused laser beam that is detected in the Fourier plane with a split detector. The signal from the split detector is differenced, which plays a role in the electronic domain similar to that of a phase plate in optical phase contrast imaging. The PC-Class BioCD is simple in both theory and implementation, requiring no microstructure fabrication and no complex detection. Its potential in high speed label-free biosensing is demonstrated by a two-analyte immunoassay that shows good rejection of nonspecific binding and low antibody cross-reactivity. Immunoassays were performed against IgG immunoglobulins with detection of bound analyte on pictogram level. To show the potential of scaling up to hundreds or thousands of analytes per disk, an experiment was also performed with small drops of protein solution.
机译：我们之前曾介绍过生物光盘（BioCD）作为灵敏的检测平台，用于检测固定在旋转磁盘表面上的图案化生物分子。旋转盘干涉仪可以高速检测（每点10微秒）光程长度变化，直至亚纳米级，并具有很高的可重复性。在机械旋转的磁盘上执行稳定干涉测量的关键是自参考：独立于机械振动或相对运动，将信号和参考光束的相位锁定为正交（π/ 2相差）。以前已经报道了BioCD的两个正交类别：微衍射类别（MD-Class）和自适应光学类别（AO-Class）{Peng，2004＃565; Varma，2004＃440}。在本文中，我们介绍了第三类BioCD，即相差类（PC类）BioCD。使用光刻法以1024辐条图案将蛋白质固定在磁盘上。印制的蛋白质图案的边缘会衍射聚焦的激光束，该激光束在傅立叶平面中使用分离检测器检测到。来自分离检测器的信号是不同的，其在电子域中的作用类似于光学相衬成像中的相板。 PC级BioCD在理论和实施上都很简单，不需要微结构的制造，也不需要复杂的检测。两种分析物的免疫分析法显示了其在无标记生物高速传感中的潜力，该分析法显示出对非特异性结合的良好排斥和较低的抗体交叉反应性。针对IgG免疫球蛋白进行免疫测定，并在象形图水平上检测结合的分析物。为了显示每个磁盘最多可扩展成百上千种分析物的潜力，还对一小滴蛋白质溶液进行了实验。

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