Skip to main content

Patch-seq: Multimodal Profiling of Single-Cell Morphology, Electrophysiology, and Gene Expression

  • Protocol
  • First Online:
Single Cell Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2752))

  • 647 Accesses

Abstract

Cells exhibit diverse morphologic phenotypes, biophysical and functional properties, and gene expression patterns. Understanding how these features are interrelated at the level of single cells has been challenging due to the lack of techniques for multimodal profiling of individual cells. We recently developed Patch-seq, a technique that combines whole-cell patch clamp recording, immunohistochemistry, and single-cell RNA-sequencing (scRNA-seq) to comprehensively profile single cells. Here we present a detailed step-by-step protocol for obtaining high-quality morphological, electrophysiological, and transcriptomic data from single cells. Patch-seq enables researchers to explore the rich, multidimensional phenotypic variability among cells and to directly correlate gene expression with phenotype at the level of single cells.

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 149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 199.99
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. Picelli S, Bjorklund AK, Faridani OR, Sagasser S, Winberg G, Sandberg R (2013) Smart-seq2 for sensitive full-length transcriptome profiling in single cells. Nat Methods 10(11):1096–1098. https://doi.org/10.1038/nmeth.2639

    Article  CAS  PubMed  Google Scholar 

  2. Tang F, Barbacioru C, Wang Y, Nordman E, Lee C, Xu N, Wang X, Bodeau J, Tuch BB, Siddiqui A, Lao K, Surani MA (2009) mRNA-Seq whole-transcriptome analysis of a single cell. Nat Methods 6(5):377–382. https://doi.org/10.1038/nmeth.1315

    Article  CAS  PubMed  Google Scholar 

  3. Eberwine J, Yeh H, Miyashiro K, Cao Y, Nair S, Finnell R, Zettel M, Coleman P (1992) Analysis of gene expression in single live neurons. Proc Natl Acad Sci U S A 89(7):3010–3014. https://doi.org/10.1073/pnas.89.7.3010

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Sucher NJ, Deitcher DL (1995) PCR and patch-clamp analysis of single neurons. Neuron 14(6):1095–1100. https://doi.org/10.1016/0896-6273(95)90257-0

    Article  CAS  PubMed  Google Scholar 

  5. Sucher NJ, Deitcher DL, Baro DJ, Warrick RM, Guenther E (2000) Genes and channels: patch/voltage-clamp analysis and single-cell RT-PCR. Cell Tissue Res 302(3):295–307. https://doi.org/10.1007/s004410000289

    Article  CAS  PubMed  Google Scholar 

  6. Subkhankulova T, Yano K, Robinson HP, Livesey FJ (2010) Grouping and classifying electrophysiologically-defined classes of neocortical neurons by single cell, whole-genome expression profiling. Front Mol Neurosci 3:10. https://doi.org/10.3389/fnmol.2010.00010

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Qiu S, Luo S, Evgrafov O, Li R, Schroth GP, Levitt P, Knowles JA, Wang K (2012) Single-neuron RNA-Seq: technical feasibility and reproducibility. Front Genet 3:124. https://doi.org/10.3389/fgene.2012.00124

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Cadwell CR, Palasantza A, Jiang X, Berens P, Deng Q, Yilmaz M, Reimer J, Shen S, Bethge M, Tolias KF, Sandberg R, Tolias AS (2016) Electrophysiological, transcriptomic and morphologic profiling of single neurons using patch-seq. Nat Biotechnol 34(2):199–203. https://doi.org/10.1038/nbt.3445

    Article  CAS  PubMed  Google Scholar 

  9. Fuzik J, Zeisel A, Mate Z, Calvigioni D, Yanagawa Y, Szabo G, Linnarsson S, Harkany T (2016) Integration of electrophysiological recordings with single-cell RNA-seq data identifies neuronal subtypes. Nat Biotechnol 34(2):175–183. https://doi.org/10.1038/nbt.3443

    Article  CAS  PubMed  Google Scholar 

  10. Cadwell CR, Scala F, Li S, Livrizzi G, Shen S, Sandberg R, Jiang X, Tolias AS (2017) Multimodal profiling of single-cell morphology, electrophysiology, and gene expression using Patch-seq. Nat Protoc 12(12):2531–2553. https://doi.org/10.1038/nprot.2017.120

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Scala F, Kobak D, Bernabucci M, Bernaerts Y, Cadwell CR, Castro JR, Hartmanis L, Jiang X, Laturnus S, Miranda E, Mulherkar S, Tan ZH, Yao Z, Zeng H, Sandberg R, Berens P, Tolias AS (2021) Phenotypic variation of transcriptomic cell types in mouse motor cortex. Nature 598(7879):144–150. https://doi.org/10.1038/s41586-020-2907-3

    Article  CAS  PubMed  Google Scholar 

  12. Scala F, Kobak D, Shan S, Bernaerts Y, Laturnus S, Cadwell CR, Hartmanis L, Froudarakis E, Castro JR, Tan ZH, Papadopoulos S, Patel SS, Sandberg R, Berens P, Jiang X, Tolias AS (2019) Layer 4 of mouse neocortex differs in cell types and circuit organization between sensory areas. Nat Commun 10(1):4174. https://doi.org/10.1038/s41467-019-12058-z

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Picelli S, Bjorklund AK, Reinius B, Sagasser S, Winberg G, Sandberg R (2014) Tn5 transposase and tagmentation procedures for massively scaled sequencing projects. Genome Res 24(12):2033–2040. https://doi.org/10.1101/gr.177881.114

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Jiang X, Shen S, Cadwell CR, Berens P, Sinz F, Ecker AS, Patel S, Tolias AS (2015) Principles of connectivity among morphologically defined cell types in adult neocortex. Science 350(6264):aac9462. https://doi.org/10.1126/science.aac9462

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Kobak D, Berens P (2019) The art of using t-SNE for single-cell transcriptomics. Nat Commun 10(1):5416. https://doi.org/10.1038/s41467-019-13056-x

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Kobak D, Bernaerts Y, Weis MA, Scala F, Tolias AS, Berens P (2021) Sparse reduced-rank regression for exploratory visualisation of paired multivariate data. J R Stat Soc Ser C Appl Stat 70(4):980–1000. https://doi.org/10.1111/rssc.12494

    Article  Google Scholar 

  17. Tripathy SJ, Toker L, Bomkamp C, Mancarci BO, Belmadani M, Pavlidis P (2018) Assessing transcriptome quality in patch-seq datasets. Front Mol Neurosci 11:363. https://doi.org/10.3389/fnmol.2018.00363

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Cathryn R. Cadwell .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2024 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Cadwell, C.R., Tolias, A.S. (2024). Patch-seq: Multimodal Profiling of Single-Cell Morphology, Electrophysiology, and Gene Expression. In: Gužvić, M. (eds) Single Cell Analysis. Methods in Molecular Biology, vol 2752. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3621-3_15

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-3621-3_15

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3620-6

  • Online ISBN: 978-1-0716-3621-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics