| RRC ID |
69161
|
| Author |
Gebreyesus ST, Siyal AA, Kitata RB, Chen ES, Enkhbayar B, Angata T, Lin KI, Chen YJ, Tu HL.
|
| Title |
Streamlined single-cell proteomics by an integrated microfluidic chip and data-independent acquisition mass spectrometry.
|
| Journal |
Nat Commun
|
| Abstract |
Single-cell proteomics can reveal cellular phenotypic heterogeneity and cell-specific functional networks underlying biological processes. Here, we present a streamlined workflow combining microfluidic chips for all-in-one proteomic sample preparation and data-independent acquisition (DIA) mass spectrometry (MS) for proteomic analysis down to the single-cell level. The proteomics chips enable multiplexed and automated cell isolation/counting/imaging and sample processing in a single device. Combining chip-based sample handling with DIA-MS using project-specific mass spectral libraries, we profile on average ~1,500 protein groups across 20 single mammalian cells. Applying the chip-DIA workflow to profile the proteomes of adherent and non-adherent malignant cells, we cover a dynamic range of 5 orders of magnitude with good reproducibility and <16% missing values between runs. Taken together, the chip-DIA workflow offers all-in-one cell characterization, analytical sensitivity and robustness, and the option to add additional functionalities in the future, thus providing a basis for advanced single-cell proteomics applications.
|
| Volume |
13(1)
|
| Pages |
37
|
| Published |
2022-1-10
|
| DOI |
10.1038/s41467-021-27778-4
|
| PII |
10.1038/s41467-021-27778-4
|
| PMID |
35013269
|
| PMC |
PMC8748772
|
| MeSH |
Animals
Cell Line, Tumor
Cell Separation
Humans
Lab-On-A-Chip Devices*
Lung Neoplasms
Mass Spectrometry / methods*
Microfluidics / methods*
Proteome
Proteomics / methods*
Reproducibility of Results
Workflow
|
| IF |
12.121
|
| Resource |
| Human and Animal Cells |
PC-9(RCB4455) |
