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Multimodal profiling of single-cell morphology, electrophysiology, and gene expression using Patch-seq.


ABSTRACT: Neurons exhibit a rich diversity of morphological phenotypes, electrophysiological properties, and gene-expression patterns. Understanding how these different characteristics are interrelated at the single-cell level has been difficult because of 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 neurons from mouse brain slices. Here, we present a detailed step-by-step protocol, including modifications to the patching mechanics and recording procedure, reagents and recipes, procedures for immunohistochemistry, and other tips to assist researchers in obtaining high-quality morphological, electrophysiological, and transcriptomic data from single neurons. Successful implementation of Patch-seq allows researchers to explore the multidimensional phenotypic variability among neurons and to correlate gene expression with phenotype at the level of single cells. The entire procedure can be completed in ?2 weeks through the combined efforts of a skilled electrophysiologist, molecular biologist, and biostatistician.

SUBMITTER: Cadwell CR 

PROVIDER: S-EPMC6422019 | biostudies-literature | 2017 Dec

REPOSITORIES: biostudies-literature

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Multimodal profiling of single-cell morphology, electrophysiology, and gene expression using Patch-seq.

Cadwell Cathryn R CR   Scala Federico F   Li Shuang S   Livrizzi Giulia G   Shen Shan S   Sandberg Rickard R   Jiang Xiaolong X   Tolias Andreas S AS  

Nature protocols 20171116 12


Neurons exhibit a rich diversity of morphological phenotypes, electrophysiological properties, and gene-expression patterns. Understanding how these different characteristics are interrelated at the single-cell level has been difficult because of 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  ...[more]

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