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ORIGINAL RESEARCH article

Front. Chem.
Sec. Analytical Chemistry
Volume 12 - 2024 | doi: 10.3389/fchem.2024.1428547

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

Provisionally accepted
Stanislau Stanisheuski Stanislau Stanisheuski 1Arpa Ebrahimi Arpa Ebrahimi 1Kavi A. Vaidya Kavi A. Vaidya 1Hyo Sang Jang Hyo Sang Jang 2Liping Yang Liping Yang 1Alex J. Eddins Alex J. Eddins 1Carrie Marean-Reardon Carrie Marean-Reardon 1Maria Clara Franco Maria Clara Franco 3Claudia S. Maier Claudia S. Maier 1*
  • 1 Oregon State University, Corvallis, United States
  • 2 Hewlett-Packard (United States), Palo Alto, California, United States
  • 3 Florida International University, Miami, Florida, United States

The final, formatted version of the article will be published soon.

    In this study, we adapted an HP D100 Single Cell Dispensera novel low-cost thermal inkjet (TIJ) platform with impedance-based single cell detectionfor dispensing of individual cells and one-pot sample preparation. We repeatedly achieved label-free identification of up to 1300 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.

    Keywords: Single-cell proteomics, Thermal inkjet, single-cell dispensing, label-free, Automation, Accessible, Commercially available

    Received: 06 May 2024; Accepted: 31 Jul 2024.

    Copyright: © 2024 Stanisheuski, Ebrahimi, Vaidya, Jang, Yang, Eddins, Marean-Reardon, Franco and Maier. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Claudia S. Maier, Oregon State University, Corvallis, United States

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.