Zero-mode waveguide device with microlens for enhancing fluorescence signal intensity

被引：0
作者：
Fu B.-W. [1 ,2 ]
Guo Z. [2 ,3 ,4 ]
Yu P.-F. [2 ,5 ]
Li C.-Y. [2 ,6 ]
Zhang Z.-Q. [2 ,5 ]
Zhou L.-Q. [1 ,2 ]
机构：
[1] Academy of Engineering & Technology, Fudan University, Shanghai
[2] Key Laboratory of Biomedical Detection Technology, Suzhou
[3] Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou
[4] Ji Hua Laboratory, Foshan
[5] University of Science and Technology of China, Hefei
[6] Suzhou CASENS Co., Ltd, Suzhou
来源：
Guangxue Jingmi Gongcheng/Optics and Precision Engineering | 2021年 / 29卷 / 08期
关键词：
Fluorescence signal detection; Microlens; Nanopore; Zero mode waveguides;
D O I：
10.37188/OPE.20212908.1921
中图分类号：
学科分类号：
摘要：
In this study, an improved zero-mode waveguide device is designed and developed. This device is used to enhance fluorescence signal intensity and eliminate fluorescence crosstalk. The device is fabricated using micro/nanofabrication technologies. It reduces the divergence angle of fluorescence through a combination of a microlens and nanopores; the constructive interference of fluorescence enhances the intensity of the detection signal. Specifically, electron beam lithography is used to prepare a nanohole array with a controllable diameter, and a microlens array with an adjustable angle is fabricated by combining ultraviolet lithography and reactive ion etching. ImageJ is used to read the fluorescence gray value for signal comparison. Compared with a standard zero-film waveguide device, the improved device increases the fluorescence signal intensity by 14.5 times and the signal-to-noise ratio by 9 times. As determined through SEM characterization, the nanopore diameter of the device is (100.3 ± 4.9) nm, and the inclination angle of the microlens is (21.1 ± 0.7)°. The improved zero-mode waveguide device can effectively mitigate fluorescence crosstalk while enhancing fluorescence signal intensity. Related research provides a feasible solution to the problems of weak fluorescence detection and fluorescence crosstalk. © 2021, Science Press. All right reserved.
引用
收藏
页码：1921 / 1930
页数：9
相关论文
共 26 条
  • [11] PATRA S, BAIBAKOV M, CLAUDE J B, Et al., Surface passivation of zero-mode waveguide nanostructures: benchmarking protocols and fluorescent labels, Scientific reports, 10, 1, pp. 1-10, (2020)
  • [12] GOLDSCHEN-OHM M P, WHITE D S, KLENCHIN V A, Et al., Observing single-molecule dynamics at millimolar concentrations, Angewandte Chemie, 129, 9, pp. 2439-2442, (2017)
  • [13] BAIBAKOV M, PATRA S, CLAUDE J B, Et al., Extending single-molecule förster resonance energy transfer (FRET) range beyond 10 nanometers in zero-mode waveguides, ACS nano, 13, 7, pp. 8469-8480, (2019)
  • [14] ZAMBRANA-PUYALTO X, PONZELLINI P, MACCAFERRI N, Et al., A hybrid metal-dielectric zero mode waveguide for enhanced single molecule detection, Chemical Communications, 55, 65, pp. 9725-9728, (2019)
  • [15] FU K, KWON S R, HAN D, Et al., Single entity electrochemistry in nanopore electrode arrays: ion transport meets electron transfer in confined geometries, Accounts of chemical research, 53, 4, pp. 719-728, (2020)
  • [16] LEVENE M J, KORLACH J, TURNER S W, Et al., Zero-mode waveguides for single-molecule analysis at high concentrations, science, 299, 5607, pp. 682-686, (2003)
  • [17] ZHOU P, WANG Z H, HAN X L, Et al., Adhesive Mounting Deformation of Ultra-thin Quartz Workpiece in Precision Machining [J], Opt. Precision Eng, 27, 11, pp. 2402-2409, (2019)
  • [18] ZHAO J, BRANAGAN S P, BOHN P W., Single-molecule enzyme dynamics of monomeric sarcosine oxidase in a gold-based zero-mode waveguide, Applied spectroscopy, 66, 2, pp. 163-169, (2012)
  • [19] CASTNER D G, RATNER B D., Biomedical surface science: Foundations to frontiers, Surface Science, 500, 1-3, pp. 28-60, (2002)
  • [20] SAMIEE K T, FOQUET M, GUO L, Et al., λ-repressor oligomerization kinetics at high concentrations using fluorescence correlation spectroscopy in zero-mode waveguides, Biophysical journal, 88, 3, pp. 2145-2153, (2005)
123