| RRC ID |
89521
|
| Author |
Kamarulzaman L, Kim S, Hidaka T, Tsuchida M, Taniguchi Y.
|
| Title |
Molecular profiling of the single-cell proteome via gel electrophoresis and 3D single-molecule imaging.
|
| Journal |
Nat Commun
|
| Abstract |
Recent advances in shotgun proteomics and immunoassays have yielded powerful single-cell proteomics technologies. However, current methods lack the sensitivity required to comprehensively quantify protein abundances in individual cells. Here, we present single-cell PAGE-PISA, an ultra-sensitive proteome profiling strategy that combines gel electrophoresis with 3D single-molecule fluorescence imaging. Our approach labels all proteins in single cells with fluorescent dyes, separates them by electrophoresis, and counts with single-molecule resolution. This technique quantified over 107 protein copies from a single mammalian cell with the sensitivity to detect low-abundance proteins down to 105 copies per species. Single-cell PAGE-PISA successfully classified cells into distinct cell types based on their proteomic profiles. Furthermore, our single-cell proteome data strongly correlated with predicted developmental states during cardiomyocyte differentiation, providing complementary information to single-cell transcriptome data. Together, single-cell PAGE-PISA enables highly sensitive and quantitative proteome profiling at the single-cell level, capturing subtle proteomic differences that distinguish diverse cellular states.
|
| Volume |
17(1)
|
| Published |
2026-7-8
|
| DOI |
10.1038/s41467-026-74840-0
|
| PII |
10.1038/s41467-026-74840-0
|
| PMID |
42420286
|
| PMC |
PMC13346896
|
| MeSH |
Animals
Cell Differentiation
Electrophoresis, Polyacrylamide Gel* / methods
Fluorescent Dyes
Myocytes, Cardiac / cytology
Myocytes, Cardiac / metabolism
Proteome* / metabolism
Proteomics* / methods
Single Molecule Imaging* / methods
Single-Cell Analysis* / methods
|
| Resource |
| Human and Animal Cells |
HeLa(RCB0007)
PC-3(RCB2145) |
