高信噪比荧光检测微反射器零模波导器件的研制

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中图分类号：TP212. 3 文献标识码：A

doi：10. 37188/OPE. 20253307. 1091 CSTR：32169. 14. OPE. 20253307. 1091

Development of zero-mode waveguide device with micro-reflector for high signal-to-noise fluorescence detection

LIU Xuanshuo 1，2 ，LI Chuanyu 1，2 ，JIAO Chunxiao 1，2 ，HUANG Runhu 1，2 ， ZHOU Lianqun 1，2 *，GUO Zhen 1，2. *

（1. University of Science and Technology of China，Hefei ，China； 2. Suzhou Institute of Biomedical Engineering and Technology， Chinese Academy of Sciences，Suzhou ，China） * Corresponding author， E -mail：zhoulq@sibet. ac. cn；guozhen@sibet. ac. cn

Abstract：This study addresses the limitations of low signal collection efficiency and suboptimal signal-tonoise ratio in zero-mode waveguide（ZMW）devices employed for fluorescence detection. A micro-reflec⁃ tor ZMW device was designed and fabricated to enhance the signal-to-noise ratio in fluorescence measure⁃ ments. Finite element electromagnetic simulations demonstrated a significant improvement in the spatial distribution and intensity of excitation and emission fields within the device. The ZMW apertures and a three-stage micro-reflector structure ，with a height of 6.48μm and a bottom diameter of 4.24μm ，were fabricated using a thick adhesive process followed by exposure etching ，complemented by focused ion beam perforation and additional micro-nano fabrication techniques. Fluorescence performance was evalu⁃ ated using fluorescent microspheres，and detection efficiency was assessed through fluorescence imaging. Quantitative analysis of grayscale intensity and signal-to-noise ratio revealed that the developed device achieved a fluorescence detection signal-to-noise ratio of 119. 6，representing an approximate 4. 27-fold enhancement compared to conventional ZMW devices without the micro-reflector. This high signal-tonoise ratio ZMW device demonstrates significant potential for applications in weak fluorescence detection and related fields.

Key words：zero-mode waveguide；micro-reflector；signal-to-noise ratio；fluorescence signal detection

1 引 言

高灵敏度光学检测在分子生物学和细胞生物学等领域至关重要［1-3］，目前通常采用全内反射［4-5］等检测方法。（剩余10803字）