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

in Frontiers in chemistry by Stanislau Stanisheuski, Arpa Ebrahimi, Kavi Aashish Vaidya, Hyo Sang Jang, Liping Yang, Alex Jordan Eddins, Carrie Marean-Reardon, Maria Clara Franco, Claudia Susanne Maier

TLDR

  • The study developed a novel low-cost thermal inkjet (TIJ) platform for dispensing individual cells and one-pot sample preparation, achieving label-free identification of up to 1,300 proteins from a single cell in a single run.

Abstract

In this study, we adapted an HP D100 Single Cell Dispenser - a novel low-cost thermal inkjet (TIJ) platform with impedance-based single cell detection - for dispensing of individual cells and one-pot sample preparation. We repeatedly achieved label-free identification of up to 1,300 proteins from a single cell in a single run using an Orbitrap Fusion Lumos Mass Spectrometer coupled to either an Acquity UPLC M-class system or a Vanquish Neo UHPLC system. The developed sample processing workflow is highly reproducible, robust, and applicable to standardized 384- and 1536-well microplates, as well as glass LC vials. We demonstrate the applicability of the method for proteomics of single cells from multiple cell lines, mixed cell suspensions, and glioblastoma tumor spheroids. As additional proof of robustness, we monitored the results of genetic manipulations and the expression of engineered proteins in individual cells. Our cost-effective and robust single-cell proteomics workflow can be transferred to other labs interested in studying cells at the individual cell level.

Overview

  • The study developed a novel low-cost thermal inkjet (TIJ) platform for dispensing individual cells and one-pot sample preparation.
  • A high-performance liquid chromatography (HPLC) system was coupled to a mass spectrometer to detect up to 1,300 proteins from a single cell in a single run.
  • The developed sample processing workflow is highly reproducible, robust, and applicable to standardized 384- and 1536-well microplates, as well as glass LC vials.

Comparative Analysis & Findings

  • The study achieved label-free identification of up to 1,300 proteins from a single cell in a single run using an Orbitrap Fusion Lumos Mass Spectrometer.
  • The developed sample processing workflow is highly reproducible and robust.
  • The method is applicable to standardized 384- and 1536-well microplates, as well as glass LC vials.

Implications and Future Directions

  • This cost-effective and robust single-cell proteomics workflow can be transferred to other labs interested in studying cells at the individual cell level.
  • Future studies can focus on optimizing the method for specific cell types or applications.
  • The method can be used to study genomics, transcriptomics, and other -omics level analysis of individual cells.
Read Full Article