Simultaneous Targeted Detection of Proteins and RNAs in Single Cells

Aik T Ooi; David W Ruff

doi:10.1007/978-1-4939-9240-9_22

Simultaneous Targeted Detection of Proteins and RNAs in Single Cells

Methods Mol Biol. 2019:1979:379-392. doi: 10.1007/978-1-4939-9240-9_22.

Authors

Aik T Ooi^{1

2}, David W Ruff³

Affiliations

¹ Fluidigm Corporation, South San Francisco, CA, USA. ooi@missionbio.com.
² Mission Bio, Inc., South San Francisco, CA, USA. ooi@missionbio.com.
³ Mission Bio, Inc., South San Francisco, CA, USA.

PMID: 31028649
DOI: 10.1007/978-1-4939-9240-9_22

Abstract

Simultaneous detection of both RNA and protein in individual single cells offers a powerful tool for genotype-to-phenotype investigations. Proximity extension assay (PEA) is a quantitative, sensitive, and multiplex protein detection system that has superb utility in single-cell omic analysis. We implemented PEA using the flexible microfluidic workflow of the Fluidigm^® C1™ system followed by real-time quantitative polymerase chain reaction (RT-qPCR) on the Fluidigm Biomark™ HD system. With this workflow, targeted quantification of RNAs and proteins within individual cells is readily conducted.

Keywords: Antibody detection; Microfluidics; Multi-omics; Protein quantification; Protein–RNA correlation; Proximity extension assay (PEA); RT-qPCR; Single-cell analysis; Single-cell protein detection; qPCR.

MeSH terms

Animals
Gene Expression Profiling / methods*
Humans
Microfluidic Analytical Techniques / methods*
Proteins / analysis*
Proteins / genetics
RNA / analysis*
RNA / genetics
Real-Time Polymerase Chain Reaction / methods
Single-Cell Analysis / methods*
Workflow

Substances

Proteins
RNA