摘要
数字聚合酶链式反应(dPCR)是一种能实现绝对定量检测的分子生物学技术。传统的dPCR荧光检测系统在检测通量、检测速度、系统体积以及成本之间难以达到合理平衡。特别是传统照明系统设计在照射面积、均匀性、体积和开关响应速度方面存在限制,影响设备的整体性能及应用。为此,本文设计了一种基于匀光设计LED激发优化的便携式dPCR大视场荧光检测系统,旨在不牺牲检测速度与通量的前提下,缩小系统体积并降低成本。系统通过高集成度的环形LED光源排布,实现了18mm×18mm全视场范围内三种荧光通道(FAM、HEX、ROX)的均匀照明,实际照明均匀度超过90%,并支持三通道快速开关切换。相比于传统的照明模块,LED匀光激发照明在小型化、低成本以及高集成度方面具有优势,适应于多种荧光检测应用场景,促进dPCR技术未来的发展。
Digital Polymerase Chain Reaction(dPCR)is a molecular biology technique that enables absolute quantifica-tion.However,conventional dPCR fluorescence detection systems face challenges in achieving an optimal balance among de-tection throughput,speed,system size,and cost.In particular,traditional illumination system designs are constrained by limitations in illumination area,uniformity,volume,and switching response,which affect the overall performance and appli-cability of the equipment.To address these issues,this study proposes a portable dPCR fluorescence detection system opti-mized for large-field imaging based on an LED excitation design with uniform illumination.This system aims to reduce size and costs without compromising detection speed and throughput.Utilizing a high-integration ring-shaped LED light source ar-rangement,the system achieves uniform illumination across an 18 mm×18 mm field of view for three fluorescence channels(FAM,HEX,ROX),with actual illumination uniformity exceeding 90%,and supports rapid three-channel switching.Compared with traditional illumination modules,the LED-based uniform excitation illumination offers advantages in minia-turization,cost-efficiency,and integration,making it suitable for various fluorescence detection applications and advancing the future development of dPCR technology.
作者
金旭东
张磬瀚
严言
郑继红
韦晓孝
万新军
JING Xudong;ZHANG Qinhan;YAN Yan;ZHENG Jihong;WEI Xiaoxiao;WAN Xinjun(School of Optical-Electrical and Computer Engineering,University of Shanghai for Science and Technology,Shanghai 200093,China;Shanghai Key Lab of Modern Optical System,University of Shanghai for Science and Technology,Shanghai 200093,China)
出处
《光学技术》
北大核心
2025年第4期424-431,共8页
Optical Technique
基金
上海市优秀学术/技术带头人计划(22XD1401000)。
关键词
应用光学
数字PCR
微流控芯片
光学设计
荧光显微镜
applied optics
digital PCR technology
microfluidic chip
optical design
fluorescence microscopy
