Design of the fluorescent LED illumination in a Wide-field dPCR detection system

JING Xudong; ZHANG Qinhan; YAN Yan; ZHENG Jihong; WEI Xiaoxiao; WAN Xinjun

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Design of the fluorescent LED illumination in a Wide-field dPCR detection system

Optical Devices & System Design | 更新时间：2025-10-09

- Design of the fluorescent LED illumination in a Wide-field dPCR detection system
- OPTICAL TECHNIQUE Vol. 51, Issue 4, Pages: 424-431(2025)
- 作者机构：
  
  1.上海理工大学光电信息与计算机学院，上海 200093
  2.上海理工大学上海市现代光学系统重点实验室，上海 200093
- 作者简介：
- 基金信息：
- DOI：
  CLC： TH742
- Received：19 November 2024，
  
  Revised：2025-02-27，
  
  Published：20 July 2025
- 稿件说明：
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金旭东,张磬瀚,严言等.大视场dPCR检测系统中的荧光LED照明设计[J].光学技术,2025,51(04):424-431.

JING Xudong,ZHANG Qinhan,YAN Yan,et al.Design of the fluorescent LED illumination in a Wide-field dPCR detection system[J].OPTICAL TECHNIQUE,2025,51(04):424-431.
金旭东,张磬瀚,严言等.大视场dPCR检测系统中的荧光LED照明设计[J].光学技术,2025,51(04):424-431. DOI：

JING Xudong,ZHANG Qinhan,YAN Yan,et al.Design of the fluorescent LED illumination in a Wide-field dPCR detection system[J].OPTICAL TECHNIQUE,2025,51(04):424-431. DOI：

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。

Abstract

Digital Polymerase Chain Reaction （dPCR） is a molecular biology technique that enables absolute quantification. However

conventional dPCR fluorescence detection systems face challenges in achieving an optimal balance among detection 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 applicability of the equipment. To address these issues

this study proposes a portable dPCR fluorescence detection system optimized 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 arrangement

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 miniaturization

cost-efficiency

and integration

making it suitable for various fluorescence detection applications and advancing the future development of dPCR technology.

关键词

Keywords

references

Vogelstein B , Kinzler K W . Digital PCR [J ] . Proceedings of the National Academy of Sciences , 1999 , 96 （ 16 ）: 9236 — 9241 .

Bartlett J M S , Stirling D . A short history of the polymerase chain reaction [J ] . Methods in Molecular Biology （Clifton, NJ） , 2003 , 226 : 3 — 6 .

Quan P L , Sauzade M , Brouzes E . dPCR: A technology review [J ] . Sensors , 2018 , 18 （ 4 ）: 1271 .

Taylor S C , Laperriere G , Germain H . Droplet digital PCR versus qPCR for gene expression analysis with low abundant targets: From variable nonsense to publication quality data [J ] . Scientific Reports , 2017 , 7 （ 1 ）: 2409 .

Ren M S , Lin H , Chen S J , et al . Detection of pseudorabies virus by duplex droplet digital PCR assay [J ] . Journal of Veterinary Diagnostic Investigation , 2018 , 30 （ 1 ）: 105 — 112 .

Niu M C , Cao W J , Wang Y C , et al . Droplet-based transcriptome profiling of individual synapses [J ] . Nature Biotechnology , 2023 , 41 （ 9 ）: 1332 .

Takeshita T , Iwase H . dPCR mutational analyses in cell-free DNA: A comparison with tissues [J ] . Methods in Molecular Biology , 2019 , 1909 : 105 — 118 .

Yi K , Wang X , Filippov S K , Zhang H . Emerging ctDNA detection strategies in clinical cancer theranostics [J ] . Smart Medicine , 2023 , 2 （ 4 ）: e20230031 .

Chin N A , Salihah N T , Shivanand P , Ahmed M U . Recent trends and developments of PCR-based methods for the detection of food-borne Salmonella bacteria and Norovirus [J ] . Journal of Food Science and Technology , 2022 , 59 （ 12 ）: 4570 — 4582 .

Ottesen E A , Hong J W , Quake S R , Leadbetter J R . Microfluidic digital PCR enables multigene analysis of individual environmental bacteria [J ] . Science , 2006 , 314 （ 5804 ）: 1464 — 1467 .

Morneau D . Droplet-based microfluidics [J ] . Nature Reviews Methods Primers , 2023 , 3 （ 1 ）: 32

Zhang P F , Abate A R . High-definition single-cell printing: Cell-by-cell fabrication of biological structures [J ] . Advanced Materials , 2020 , 32 （ 52 ）: e2005346 .

Beer N R , Wheeler E K , Lee-Houghton L , et al . On-chip single-copy real-time reverse-transcription PCR in isolated picoliter droplets [J ] . Analytical Chemistry , 2008 , 80 （ 6 ）: 1854 — 1858 .

Clark I C , Mudvari P , Thaploo S , et al . HIV silencing and cell survival signatures in infected T cell reservoirs [J ] . Nature , 2023 , 614 （ 7947 ）: 318 — 325 .

Sun A C , Steyer D J , Allen A R , et al . A droplet microfluidic platform for high-throughput photochemical reaction discovery [J ] . Nature Communications , 2020 , 11 （ 1 ）: 6202 .

Jacobs B K , Goetghebeur E , Clement L . Impact of variance components on reliability of absolute quantification using digital PCR [J ] . BMC Bioinformatics , 2014 , 15 （ 1 ）: 283 .

Sidstedt M , Romsos EL , Hedell R , et al . Accurate digital polymorase chain reaction quantification of challenging samples applying inhibitor-toberant DNA pdymerases [J ] . Analytical Chemistry , 2017 , 89 （ 3 ）: 1642 — 1649 .

鲍猛 , 张磬瀚 , 郑继红 , 等 . 基于波前编码的大像面荧光显微成像系统景深延拓研究 [J ] . 光学技术 , 2024 , 50 （ 04 ）: 411 — 418 .

Bao Meng , Zhang Qinghan , Zheng Jihong , et al . [J]. Continuation of the depth of field of large-area fluorescence microscopic imaging system based on wavefront coding [J ] . Optical technology , 2024 , 50 （ 04 ）: 411 — 418 .

朱天赟 , 郑继红 , 孙刘杰 , 等 . 基于图像拼接的高通量数字PCR荧光基因芯片读取系统的设计 [J ] . 光学技术 , 2019 , 45 （ 01 ）: 107 — 111 .

Zhu Tianyun ， Zheng Jihong ， Sun Liujie , et al . Design of high throughput digital PCR fluorescent gene chip detection system based on image mosaic [J ] . Optical technology , 2019 , 45 （ 1 ）: 107 — 111

李斌 , 唐金龙 , 于文豪 , 等 . 拼接误差对拼接镜成像质量的影响 [J ] . 激光与光电子学进展 , 2017 , 54 （ 07 ）: 185 — 194 .

Li , Bin , Tang , Jinlong , Yu , Wenhao , et al . Effects of Stitching Errors on the Imaging Quality of Stitched Mirrors [J ] . Laser & Optoelectronics Progress , 2017 , 54 （ 07 ）: 185 — 194 .

Zhou S , Gou T , Hu J , et al . A highly integrated real-time digital PCR device for accurate DNA quantitative analysis [J ] . Biosensors and Bioelectronics , 2019 , 128 : 151 — 158 .

苟彤 . 基于智能手机的手持式数字PCR系统开发及其应用研究 [D ] , 浙江大学 , 2019 .

Gou Tong . Development and application research of a handheld digital PCR system based on smartphones [D ] , Zhejiang University , 2019 .

张鑫 . 高通量基因测序荧光显微成像光学系统研究 [D ] . 中国科学院大学 , 2018 .

Zhang Xin . Research on optical system for fluorescence microscopy imaging in High-throughput gene sequencing [D ] , University of Chinese Academy of Sciences 2018 .

刘美辰 , 孟庆宇 , 张洪文 , 等 . 双波段LED照明的基因测序荧光显微系统设计 [J ] . 光学精密工程 , 2024 , 32 （ 16 ）: 2455 — 2463 .

Liu M , Meng Q , Zhang H , et al . Design of gene sequencing fluorescence microscopic system with dual-band LED illumination [J ] . Opt Precis Eng （China） , 2024 , 32 （ 16 ）: 2455 — 2463 .

Koo S B N , Kim Y S , Park C Y , Lee D J . Compact camera fluorescence detector for parallel-light lens-based real-time PCR system [J ] . Sensors , 2022 , 22 （ 21 ）: 8575 .

于岩峰 . 基于免疫荧光定量检测系统的研究及激光光源优化 [D ] ; 山东师范大学 , 2024 .

Yu Yanfeng . Researoh of immunoflaorescence-based quautitative detection system and optimization of Laser source [D ] . Shandong Normal University , 2024 .

Zi J , Bi H . Fluorescence microscope light source based on integrated LED [J ] . Light: Science & Applications , 2023 , 12 （ 1 ）: 224 .

Li C , Kang N , Ye S , et al . All-in-one OsciDrop digital PCR system for automated and highly multiplexed molecular diagnostics [J ] . Advanced Science , 2024 , 11 （ 21 ）: e2309557 .

Gaňová M , Wang X , Yan Z , et al . Temperature non-uniformity detection on dPCR chips and temperature sensor calibration [J ] . RSC Advances , 2022 , 12 （ 4 ）: 2375 — 2382 .

陈诚 , 郑继红 , 万新军 , 等 . 应用于dPCR基因芯片的荧光精缩无限远物镜设计 [J ] . 光学技术 , 2019 , 45 （ 05 ）: 518 — 525 .

Chen Cheng , Zheng Jihong , Wan Xinjun , et al . Design of fluorescent Infinite-distance contraction objective for dPCR gene chip . Optical Technique , 2019 , 45 （ 5 ）: 518 — 525 .

谢欣茹 , 杨波 , 潘帅 , 等 . 多通道PCR荧光检测仪的光学系统设计 [J ] . 光学技术 , 2019 , 45 （ 05 ）: 531 — 4+640 .

Xie Xinru , Yang Bo , Pan Shuai , et al . Optical system design of multichannel PCR fluorescence detector（Article） [J ] . Guangxue Jishu/Optical Technique , 2019 , 45 （ 5 ）: 531 — 534,640

王子程 , 郑继红 , 万新军 , 等 . 应用于dPCR的大视场荧光显微检测系统的设计 [J ] . 光学技术 , 2021 , 47 （ 01 ）: 1 — 5+22 .

Wang Zicheng , Zheng Jihong , Wan Xinjun , et al . Design of fluorescence microscopy detection system with wide field of view for dPCR [J ] . Optical Technique , 2021 , 47 （ 1 ）: 6 .

徐振超 , 张磬瀚 , 郑继红 , 等 . 大视场dPCR芯片荧光检测系统的设计与优化 [J ] . 光学技术 , 2023 , 49 （ 04 ）: 502 — 507 .

Xu Zhenchao , Zhang Qinghan , Zheng Jihong , et al . Design and optimization of fluorescence detection system for Large-Field-of-view dPCR chip [J ] . Optical Technique , 2023 , 49 （ 04 ）: 502 — 507 .

Poojar P , Oiye I E , Aggarwal K , et al . Repeatability of image quality in very low-field MRI [J ] . NMR in Biomedicine , 2024 , 37 （ 10 ）: e5198 .

Liu H , Nan L , Chen F , et al . Functions and applications of artificial intelligence in droplet microfluidics [J ] . Lab on a Chip , 2023 , 23 （ 11 ）: 2497 — 2513 .

Gardner K , Uddin M M , Tran L , et al . Deep learning detector for high precision monitoring of cell encapsulation statistics in microfluidic droplets [J ] . Lab on a Chip , 2022 , 22 （ 21 ）: 4067 — 4080 .

Nan L , Zhang H D , Weitz D A , Shum H C . Development and future of droplet microfluidics [J ] . Lab on a Chip , 2024 , 24 （ 5 ）: 1135 — 1153 .

Hess D , Dockalova V , Kokkonen P , et al . Exploring mechanism of enzyme catalysis by on-chip transient kinetics coupled with global data analysis and molecular modeling [J ] . Chem , 2021 , 7 （ 4 ）: 1066 — 1079 .

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