摘要
细胞异质性是理解胚胎发育、疾病演化等生命过程的关键,而传统群体细胞RNA测序无法解析单细胞层面的基因表达差异。单细胞RNA测序技术(scRNA-seq)虽能在单细胞分辨率下构建转录组图谱,但其面临单细胞分离捕获效率低、RNA微量操作偏差大等挑战。以微流控芯片为技术载体的scRNA-seq通过微米级流体操控系统,将单细胞分离、裂解、逆转录、扩增及测序文库构建等流程集成化,实现了高通量、低样本损耗及自动化操作,显著提升了scRNA-seq的效率与数据可靠性。本文概述了scRNA-seq的测序流程,包括单细胞的分离与捕获、RNA提取、逆转录与扩增、单细胞测序等步骤,分析了微流控芯片在适配单细胞、精准控制反应体积和实现流程自动化等方面的核心优势,并简述了代表性平台Fluidigm C1、10X Genomics Chromium及BD Rhapsody的技术原理与特点。微流控芯片技术为scRNA-seq提供了高效、精准的技术平台,未来随着芯片设计的不断优化与多组学整合分析能力的提升,我们期待其在解析复杂生物系统、揭示疾病机制乃至推动精准医疗方面发挥更为深远的作用。
Cell heterogeneity is key to understanding life processes such as embryonic development and disease evolution,while traditional bulk cell RNA sequencing cannot resolve gene expression differences at the single-cell level. Although single-cell RNA sequencing (scRNA-seq) technology can construct transcriptomic maps at single-cell resolution,it faces challenges such as low efficiency in single-cell isolation and capture,and large deviations in trace RNA manipulation. Microfluidic chip technology,through a microscale fluid manipulation system,integrates processes such as single-cell isolation,lysis,reverse transcription,amplification,and sequencing library construction,achieving high-throughput,low sample loss,and automated operations,which significantly improve the efficiency and data reliability of scRNA-seq. This paper outlines the sequencing process of scRNA-seq,including steps such as single-cell isolation and capture,RNA extraction,reverse transcription and amplification,and single-cell sequencing. It analyzes the core advantages of microfluidic chips in adapting to single cells,precisely controlling reaction volumes,and realizing process automation,and briefly describes the technical principles and characteristics of representative platforms such as Fluidigm C1,10X Genomics Chromium,and BD Rhapsody. Microfluidic chip technology provides an efficient and precise technical platform for scRNA-seq. In the future,with the continuous optimization of chip design and the improvement of multi-omics integrated analysis capabilities,we expect it to play a more profound role in resolving complex biological systems,revealing disease mechanisms,and even promoting precision medicine.
作者
束璐茜
张炎
Luxi Shu;Yan Zhang(Faculty of Science,Kunming University of Science and Technology,Kunming 650500,China)
出处
《化学进展》
北大核心
2026年第2期283-297,共15页
Progress in Chemistry
基金
国家自然科学基金项目(No.32460624)
昆明理工大学“双一流”创建联合专项(No.202201BE070001-028)
昆明理工大学第十九届“挑战杯”全国大学生课外学术科技作品竞赛培育项目暨2024年学生课外学术科技创新基金项目(No.2024ZK128)资助
云南省教育科学规划项目(No.BC23002)。
