Research Center for Analytical Instrumentation, Institute of Cyber-Systems and Control, College of Control Science and Engineering, State Key Laboratory of Industrial Control Technology, Zhejiang University, Hangzhou 310027, China.
Department of Hepatobiliary and Pancreatic Surgery, Huzhou Central Hospital, Huzhou Key Laboratory of Intelligent and Digital Precision Surgery, Department of General Surgery, Affiliated Huzhou Hospital, School of Medicine, Zhejiang University, Huzhou 313000, China.
Biosensors (Basel). 2024 Jan 24;14(2):64. doi: 10.3390/bios14020064.
Single-cell analysis provides an overwhelming strategy for revealing cellular heterogeneity and new perspectives for understanding the biological function and disease mechanism. Moreover, it promotes the basic and clinical research in many fields at a single-cell resolution. A digital polymerase chain reaction (dPCR) is an absolute quantitative analysis technology with high sensitivity and precision for DNA/RNA or protein. With the development of microfluidic technology, digital PCR has been used to achieve absolute quantification of single-cell gene expression and single-cell proteins. For single-cell specific-gene or -protein detection, digital PCR has shown great advantages. So, this review will introduce the significance and process of single-cell analysis, including single-cell isolation, single-cell lysis, and single-cell detection methods, mainly focusing on the microfluidic single-cell digital PCR technology and its biological application at a single-cell level. The challenges and opportunities for the development of single-cell digital PCR are also discussed.
单细胞分析为揭示细胞异质性提供了一种极具压倒性的策略，并为理解生物功能和疾病机制提供了新的视角。此外，它还以单细胞分辨率推动了许多领域的基础和临床研究。数字聚合酶链反应（dPCR）是一种具有高灵敏度和高精度的 DNA/RNA 或蛋白质绝对定量分析技术。随着微流控技术的发展，数字 PCR 已被用于实现单细胞基因表达和单细胞蛋白质的绝对定量。对于单细胞特异性基因或蛋白质的检测，数字 PCR 显示出了巨大的优势。因此，本综述将介绍单细胞分析的意义和过程，包括单细胞分离、单细胞裂解和单细胞检测方法，主要侧重于微流控单细胞数字 PCR 技术及其在单细胞水平上的生物学应用。还讨论了单细胞数字 PCR 发展的挑战和机遇。
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