Thermal inkjet makes label-free single-cell proteomics accessible and easy

Area of Science:

• Proteomics
• Cell Biology
• Biotechnology

Background:

• Single-cell analysis is crucial for understanding cellular heterogeneity.
• Existing methods for single-cell proteomics are often complex and low-throughput.

Purpose of the Study:

• To adapt a novel thermal inkjet (TIJ) platform for single-cell dispensing and sample preparation.
• To establish a robust and reproducible single-cell proteomics workflow.
• To demonstrate the applicability of the method across diverse biological samples.

Main Methods:

• Utilized an HP D100 Single Cell Dispenser with impedance-based detection.
• Employed an Orbitrap Fusion Lumos Mass Spectrometer coupled with UPLC/UHPLC systems.
• Developed a one-pot sample preparation workflow compatible with microplates and vials.

Main Results:

• Achieved label-free identification of up to 1,300 proteins per single cell.
• Demonstrated high reproducibility and robustness of the workflow.
• Successfully applied the method to cell lines, mixed cell suspensions, and tumor spheroids.

Conclusions:

• The developed workflow provides a cost-effective and scalable solution for single-cell proteomics.
• This method facilitates the study of cellular heterogeneity, genetic manipulations, and protein expression at the single-cell level.
• The workflow is transferable to other laboratories for broad application in cell biology research.