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Multimodal profiling of single-cell morphology, electrophysiology, and gene expression using Patch-seq.
Cathryn R Cadwell1, Federico Scala1, Shuang Li1
1Department of Neuroscience, Baylor College of Medicine, Houston, Texas, USA.
Nature Protocols
|December 1, 2017
Summary
Patch-seq enables multimodal single-cell analysis of neurons by combining patch-clamp recording, immunohistochemistry, and single-cell RNA sequencing. This technique allows researchers to link gene expression to neuronal phenotype for a deeper understanding of brain cell diversity.
Area of Science:
- Neuroscience
- Molecular Biology
- Genomics
Background:
- Neurons display diverse morphologies, electrophysiology, and gene expression.
- Characterizing these traits at the single-cell level is challenging due to limited multimodal profiling techniques.
Purpose of the Study:
- To present a detailed protocol for the Patch-seq technique.
- To enable comprehensive multimodal profiling of individual neurons.
Main Methods:
- Patch-seq integrates whole-cell patch-clamp recording, immunohistochemistry, and single-cell RNA sequencing (scRNA-seq).
- The protocol includes specific modifications for patching mechanics, recording, reagents, and immunohistochemistry.
Main Results:
- High-quality morphological, electrophysiological, and transcriptomic data can be obtained from single neurons.
- The technique facilitates exploration of multidimensional phenotypic variability among neurons.
Conclusions:
- Patch-seq provides a powerful method for correlating gene expression with neuronal phenotype at the single-cell level.
- This approach enhances understanding of neuronal diversity and brain cell function.
