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Methods for Single-Cell Isolation and Preparation

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Single-cell Sequencing and Methylation

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 1255))

Abstract

Within the last decade, single-cell analysis has revolutionized our understanding of cellular processes and heterogeneity across all disciplines of life science. As the transcriptome, genome, or epigenome of individual cells can nowadays be analyzed at low cost and in high-throughput within a few days by modern techniques, tremendous improvements in disease diagnosis on the one hand and the investigation of disease-relevant mechanisms on the other were achieved so far. This relies on the parallel development of reliable cell capturing and single-cell sequencing approaches that have paved the way for comprehensive single-cell studies. Apart from single-cell isolation methods in high-throughput, a variety of methods with distinct specializations were developed, allowing for correlation of transcriptomics with cellular parameters like electrophysiology or morphology.

For all single-cell-based approaches, accurate and reliable isolation with proper quality controls is prerequisite, whereby different options exist dependent on sample type and tissue properties. Careful consideration of an appropriate method is required to avoid incorrect or biased data that may lead to misinterpretations.

In this chapter, we will provide a broad overview of the current state of the art in matters of single-cell isolation methods mostly applied for sequencing-based downstream analysis, and their respective advantages and drawbacks. Distinct technologies will be discussed in detail addressing key parameters like sample compatibility, viability, purity, throughput, and isolation efficiency.

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Acknowledgement

Funding: This work was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)—368482240/GRK2416.

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Authors and Affiliations

  1. Division of Functional Epigenetics, Institute of Zoology (Biology II), RWTH Aachen University, Aachen, Germany

    Daniel Pensold & Geraldine Zimmer-Bensch

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  1. Daniel Pensold
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  2. Geraldine Zimmer-Bensch
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Editors and Affiliations

  1. Ruijin Hospital, Shanghai Jiao Tong University, Shanghai, Shanghai, China

    Buwei Yu

  2. Department of Anesthesiology and Perioperative Medicine, Center of Clinical Single-Cell Biomedicine, Henan Provincial People’s Hospital, Zhengzhou, University People’s Hospital, Henan University People’s Hospital, Zhengzhou, P.R., China

    Jiaqiang Zhang

  3. 2nd Affiliated Hospital, Fujian Medical University, Quanzhou, Fujian, China

    Yiming Zeng

  4. Centre for Single Cell Biomedicine, Henan Provincial People’s Hospital, Zhengzhou, Henan, China

    Li Li

  5. Zhongshan Hospital, Fudan University, Shanghai, Shanghai, China

    Xiangdong Wang

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Pensold, D., Zimmer-Bensch, G. (2020). Methods for Single-Cell Isolation and Preparation. In: Yu, B., Zhang, J., Zeng, Y., Li, L., Wang, X. (eds) Single-cell Sequencing and Methylation. Advances in Experimental Medicine and Biology, vol 1255. Springer, Singapore. https://doi.org/10.1007/978-981-15-4494-1_2

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