Skip to main content

Tissue Handling and Dissociation for Single-Cell RNA-Seq

  • Protocol
  • First Online:
Single Cell Methods

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1979))

Abstract

The starting material for all single-cell protocols is a cell suspension. The particular functions and spatial distribution of immune cells generally make them easy to isolate them from the tissues where they dwell. Here we describe tissue dissociation protocols that have been used to obtain human immune cells from lymphoid and nonlymphoid tissues to be then used as input to single-cell methods. We highlight the main factors that can influence the final quality of single-cell data, namely the stress signatures that can bias its interpretation.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. van den Brink SC, Sage F, Vértesy Á et al (2017) Single-cell sequencing reveals dissociation-induced gene expression in tissue subpopulations. Nat Methods 14:935–936

    PubMed  Google Scholar 

  2. Wu YE, Pan L, Zuo Y et al (2017) Detecting activated cell populations using single-cell RNA-seq. Neuron 96:313–329.e6

    CAS  PubMed  Google Scholar 

  3. Adam M, Potter AS, Potter SS (2017) Psychrophilic proteases dramatically reduce single-cell RNA-seq artifacts: a molecular atlas of kidney development. Development 144:3625–3632

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Gunawan M, Jardine L, Haniffa M (2016) Isolation of human skin dendritic cell subsets. Methods Mol Biol 1423:119–128

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Felipe A. Vieira Braga .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2019 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Vieira Braga, F.A., Miragaia, R.J. (2019). Tissue Handling and Dissociation for Single-Cell RNA-Seq. In: Proserpio, V. (eds) Single Cell Methods. Methods in Molecular Biology, vol 1979. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9240-9_2

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9240-9_2

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9239-3

  • Online ISBN: 978-1-4939-9240-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics