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单细胞采集和分析平台。

Platforms for Single-Cell Collection and Analysis.

机构信息

Laboratory of Gene Expression, Institute of Biotechnology CAS, Biocev, Vestec 252 50, Czech Republic.

Laboratory of Growth Regulators, Faculty of Science, Palacky University, Olomouc 783 71, Czech Republic.

出版信息

Int J Mol Sci. 2018 Mar 11;19(3):807. doi: 10.3390/ijms19030807.

Abstract

Single-cell analysis has become an established method to study cell heterogeneity and for rare cell characterization. Despite the high cost and technical constraints, applications are increasing every year in all fields of biology. Following the trend, there is a tremendous development of tools for single-cell analysis, especially in the RNA sequencing field. Every improvement increases sensitivity and throughput. Collecting a large amount of data also stimulates the development of new approaches for bioinformatic analysis and interpretation. However, the essential requirement for any analysis is the collection of single cells of high quality. The single-cell isolation must be fast, effective, and gentle to maintain the native expression profiles. Classical methods for single-cell isolation are micromanipulation, microdissection, and fluorescence-activated cell sorting (FACS). In the last decade several new and highly efficient approaches have been developed, which not just supplement but may fully replace the traditional ones. These new techniques are based on microfluidic chips, droplets, micro-well plates, and automatic collection of cells using capillaries, magnets, an electric field, or a punching probe. In this review we summarize the current methods and developments in this field. We discuss the advantages of the different commercially available platforms and their applicability, and also provide remarks on future developments.

摘要

单细胞分析已成为研究细胞异质性和稀有细胞特征的一种既定方法。尽管成本高且存在技术限制,但每年在生物学的所有领域中的应用都在不断增加。随着这一趋势,单细胞分析的工具也得到了极大的发展,尤其是在 RNA 测序领域。每一次改进都提高了灵敏度和通量。大量数据的收集也刺激了生物信息学分析和解释的新方法的发展。然而,任何分析的基本要求是收集高质量的单细胞。单细胞分离必须快速、有效且温和,以保持天然的表达谱。单细胞分离的经典方法是显微操作、显微切割和荧光激活细胞分选 (FACS)。在过去的十年中,已经开发出了几种新的、高效的方法,这些方法不仅补充了传统方法,而且可能完全取代了传统方法。这些新技术基于微流控芯片、液滴、微孔板,以及使用毛细管、磁铁、电场或穿孔探针自动收集细胞。在这篇综述中,我们总结了该领域当前的方法和发展。我们讨论了不同商业上可用平台的优点及其适用性,并对未来的发展提出了看法。

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