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Updated: Jan 29, 2026
08:38
Phospho Flow Cytometry with Fluorescent Cell Barcoding for Single Cell Signaling Analysis and Biomarker Discovery
Published on: October 4, 2018
21.8K
Combined quantification of intracellular (phospho-)proteins and transcriptomics from fixed single cells
Jan P Gerlach1,2, Jessie A G van Buggenum1, Sabine E J Tanis1
1Department of Molecular Developmental Biology, Radboud Institute for Molecular Life Sciences, Science Faculty, Radboud University, PO Box 9101, 6500 HB, Nijmegen, The Netherlands.
Scientific Reports
|February 8, 2019
Summary
Single-cell RNA and Immunodetection (RAID) enables simultaneous analysis of mRNA and proteins. This method reveals how environmental signals drive diverse cellular responses and gene expression in human keratinocytes.
Area of Science:
- Cell Biology
- Molecular Biology
- Genomics
Background:
- Environmental stimuli induce varied cellular responses.
- Understanding cellular heterogeneity requires multi-omic analysis.
- Existing methods may not capture both protein and mRNA states simultaneously.
Purpose of the Study:
- To develop a method for combined transcriptome and proteome analysis in single cells.
- To investigate cellular heterogeneity in response to environmental signals.
- To validate the new method in human keratinocytes.
Main Methods:
- Development of single-cell RNA and Immunodetection (RAID).
- Application of RAID to fixed human keratinocytes.
- Analysis of both mRNA and intracellular (phospho-)proteins.
Main Results:
- RAID successfully identified differentiation-state changes at both protein and mRNA levels.
- Differentiated keratinocytes with high phosphorylated FAK (a stem cell marker) also expressed stem cell transcripts.
- Demonstrated the ability to study mRNA and phospho-proteome simultaneously.
Conclusions:
- RAID is a powerful tool for dissecting cellular heterogeneity.
- The method allows simultaneous investigation of transcriptomic and proteomic responses.
- RAID facilitates understanding of cellular responses to environmental cues at a single-cell level.
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