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采用微光纤辅助马赫-曾德尔干涉仪的无标记原位实时 DNA 杂交动力学检测。

Label-free in-situ real-time DNA hybridization kinetics detection employing microfiber-assisted Mach-Zehnder interferometer.

机构信息

Key Laboratory of Optical Information Science and Technology, Ministry of Education, Institute of Modern Optics, Nankai University, Tianjin 300071, China.

Key Laboratory of Optical Information Science and Technology, Ministry of Education, Institute of Modern Optics, Nankai University, Tianjin 300071, China.

出版信息

Biosens Bioelectron. 2016 Jul 15;81:151-158. doi: 10.1016/j.bios.2016.02.065. Epub 2016 Feb 27.

Abstract

A label-free DNA biosensor based on microfiber-assisted Mach-Zehnder interferometer (MAMZI) for in-situ real-time DNA hybridization kinetics detection has been proposed and experimentally demonstrated. A microfiber of hundreds of microns in length is fabricated by tapering a segment of standard single-mode fiber (SMF) to construct the U-shaped microcavity between the lead-in and lead-out SMFs. Thanks to the mode field mismatching between the SMF and microfiber, the incident guided mode light would separate into two beams that respectively propagate in the air microcavity and the microfiber. Consequently, interference between different light modes would occur at the joint between the microfiber and the lead-out SMF. Experimental results indicate that owing to the participation of opening cavity modes in the modal interference process, the interferometric spectrum of our proposed microcavity sensor is highly sensitive to the variation of environmental refractive index (RI), especially for the RI range around 1.34 which is useful for most biological applications. The microfiber functionalization is achieved by stepwise modifying the microfiber with monolayer Poly-l-lysine (PLL) and single-stranded DNA (ssDNA) probes to produce the sensitive surface that could uniquely attach specific target ssDNAs. The fiber surface functionalization as well as DNA hybridization processes have been experimentally investigated for different target ssDNA solutions in real time. The interferometric transmission spectrum shows large wavelength shift for different biological phases, and a detection limit conservatively down to 0.0001pmol/μL has been acquired by employing the U-shaped microcavity of 176.88μm in length. Our proposed DNA biosensor possesses several advantages such as compact size, ease of fabrication, and strong response for DNA hybridization, which make it a promising candidate for potential applications in such rapidly expanding areas as medical diagnosis, cancer screenings, medicine examination and environmental engineering, etc.

摘要

一种基于微光纤辅助马赫-曾德尔干涉仪(MAMZI)的无标记 DNA 生物传感器,用于原位实时 DNA 杂交动力学检测,已经被提出并进行了实验验证。通过将一段标准单模光纤(SMF)缩径来制作数百微米长的微光纤,从而在引入和引出 SMF 之间构建 U 形微腔。由于 SMF 和微光纤之间的模场失配,入射导模光会分离成两束光,分别在空气中微腔和微光纤中传播。因此,微光纤和引出 SMF 之间的光模干涉会发生。实验结果表明,由于开腔模式参与到模间干涉过程中,我们提出的微腔传感器的干涉光谱对环境折射率(RI)的变化非常敏感,特别是在大多数生物应用中有用的 1.34 左右的 RI 范围内。通过逐步用单层聚-L-赖氨酸(PLL)和单链 DNA(ssDNA)探针修饰微光纤,实现了微光纤的功能化,从而产生了能够独特地附着特定目标 ssDNA 的敏感表面。实验研究了不同目标 ssDNA 溶液在不同目标 ssDNA 溶液中的实时光纤表面功能化和 DNA 杂交过程。干涉透射光谱显示出不同生物阶段的大波长移动,并且通过采用 176.88μm 长的 U 形微腔,保守地获得了检测限低至 0.0001pmol/μL。我们提出的 DNA 生物传感器具有体积小、易于制造、对 DNA 杂交反应强等优点,使其成为医疗诊断、癌症筛查、医学检查和环境工程等快速发展领域的潜在应用的有前途的候选者。

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