Zhang Yan, Dhawan Anuj, Vo-Dinh Tuan
Fitzpatrick Institute for Photonics, Duke University, Durham, NC, 27708, USA.
Department of Biomedical Engineering, Duke University, Durham, NC, 27708, USA.
Nanoscale Res Lett. 2011 Dec;6(1):18. doi: 10.1007/s11671-010-9744-5. Epub 2010 Aug 31.
This paper describes the design and fabrication of fiber-optic nanoprobes developed for optical detection in single living cells. It is critical to fabricate probes with well-controlled nanoapertures for optimized spatial resolution and optical transmission. The detection sensitivity of fiber-optic nanoprobe depends mainly on the extremely small excitation volume that is determined by the aperture sizes and penetration depths. We investigate the angle dependence of the aperture in shadow evaporation of the metal coating onto the tip wall. It was found that nanoaperture diameters of approximately 50 nm can be achieved using a 25° tilt angle. On the other hand, the aperture size is sensitive to the subtle change of the metal evaporation angle and could be blocked by irregular metal grains. Through focused ion beam (FIB) milling, optical nanoprobes with well-defined aperture size as small as 200 nm can be obtained. Finally, we illustrate the use of the nanoprobes by detecting a fluorescent species, benzo[a]pyrene tetrol (BPT), in single living cells. A quantitative estimation of the numbers of BPT molecules detected using fiber-optic nanoprobes for BPT solutions shows that the limit of detection was approximately 100 molecules.
本文描述了为在单个活细胞中进行光学检测而开发的光纤纳米探针的设计与制造。制造具有良好控制的纳米孔径的探针对于优化空间分辨率和光传输至关重要。光纤纳米探针的检测灵敏度主要取决于由孔径大小和穿透深度决定的极小激发体积。我们研究了金属涂层在尖端壁上阴影蒸发过程中孔径的角度依赖性。发现使用25°倾斜角可实现约50 nm的纳米孔径直径。另一方面,孔径大小对金属蒸发角度的细微变化敏感,并且可能被不规则的金属颗粒堵塞。通过聚焦离子束(FIB)铣削,可以获得孔径大小明确至200 nm的光学纳米探针。最后,我们通过检测单个活细胞中的荧光物质苯并[a]芘四醇(BPT)来说明纳米探针的用途。对使用光纤纳米探针检测BPT溶液时检测到的BPT分子数量的定量估计表明,检测限约为100个分子。