Multimodal profiling of single-cell morphology, electrophysiology, and gene expression using Patch-seq.
Nat Protoc
; 12(12): 2531-2553, 2017 Dec.
Article
em En
| MEDLINE
| ID: mdl-29189773
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 â¼2 weeks through the combined efforts of a skilled electrophysiologist, molecular biologist, and biostatistician.
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Imuno-Histoquímica
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Análise de Sequência de RNA
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Técnicas de Patch-Clamp
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Perfilação da Expressão Gênica
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Análise de Célula Única
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Neurônios
Limite:
Animals
Idioma:
En
Revista:
Nat Protoc
Ano de publicação:
2017
Tipo de documento:
Article
País de afiliação:
Estados Unidos