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Counting Proteins in Single Cells with Addressable Droplet Microarrays
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Nanoliter-Scale Oil-Air-Droplet Chip-Based Single Cell Proteomic Analysis
Zi-Yi Li1, Min Huang2, Xiu-Kun Wang3
1Institute of Microanalytical Systems, Chemistry Department and Innovation Center for Cell Signaling Network , Zhejiang University , Hangzhou , 310058 , China.
Analytical Chemistry
|March 20, 2018
Summary
This study introduces a nanoliter-scale oil-air-droplet (OAD) chip for enhanced single-cell proteomic analysis. The OAD chip improves analytical sensitivity and efficiency for shotgun proteomics, enabling detailed cellular heterogeneity studies.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Molecular Biology
Background:
- Cellular heterogeneity is critical for understanding biological systems.
- Single-cell proteomic analysis offers deep insights but faces sensitivity challenges.
- Traditional methods often involve sample loss and lower efficiency.
Purpose of the Study:
- To develop a nanoliter-scale oil-air-droplet (OAD) chip for complex sample pretreatment and injection.
- To enhance sensitivity and efficiency in single-cell proteomic analysis using shotgun mode.
- To overcome limitations of traditional in-tube systems for single-cell proteomics.
Main Methods:
- Utilized a nanoliter-scale oil-air-droplet (OAD) chip for sample handling.
- Implemented miniaturized stationary droplet microreaction and manipulation techniques.
- Performed multistep complex sample pretreatment and injection within nanoliter droplets.
Main Results:
- Achieved minimum sample loss and high sample injection efficiency (>99%) for single cells.
- Identified 1360, 612, 192, and 51 protein IDs from 100, 50, 10, and 1 HeLa cell(s), respectively.
- Successfully analyzed single mouse oocytes, identifying 355 proteins with improved enrichment and digestion.
Conclusions:
- The OAD chip significantly increases analytical sensitivity for single-cell proteomic analysis.
- Demonstrated superior performance over traditional in-tube systems in terms of enrichment and digestion efficiency.
- The OAD chip system is a powerful tool for studying cellular heterogeneity at the single-cell level.
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