High-throughput single cell metabolomics and cellular heterogeneity exploration by inertial microfluidics coupled with pulsed electric field-induced electrospray ionization-high resolution mass spectrometry

Disheng Feng; Hang Li; Tianrun Xu; Fujian Zheng; Chunxiu Hu; Xianzhe Shi; Guowang Xu

doi:10.1016/j.aca.2022.340116

High-throughput single cell metabolomics and cellular heterogeneity exploration by inertial microfluidics coupled with pulsed electric field-induced electrospray ionization-high resolution mass spectrometry

Anal Chim Acta. 2022 Aug 15:1221:340116. doi: 10.1016/j.aca.2022.340116. Epub 2022 Jun 25.

Authors

Disheng Feng¹, Hang Li², Tianrun Xu¹, Fujian Zheng³, Chunxiu Hu⁴, Xianzhe Shi⁵, Guowang Xu⁶

Affiliations

¹ CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China; University of Chinese Academy of Sciences, Beijing, 100049, China.
² CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China.
³ CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China; University of Chinese Academy of Sciences, Beijing, 100049, China; Liaoning Province Key Laboratory of Metabolomics, Dalian, China.
⁴ CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China; Liaoning Province Key Laboratory of Metabolomics, Dalian, China.
⁵ CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China; Liaoning Province Key Laboratory of Metabolomics, Dalian, China. Electronic address: shixianzhe@dicp.ac.cn.
⁶ CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China; University of Chinese Academy of Sciences, Beijing, 100049, China; Liaoning Province Key Laboratory of Metabolomics, Dalian, China. Electronic address: xugw@dicp.ac.cn.

PMID: 35934357
DOI: 10.1016/j.aca.2022.340116

Abstract

Single cell metabolomics can obtain the metabolic profiles of individual cells and reveal cellular heterogeneity. However, high-throughput single-cell mass spectrometry (MS) analysis under physiological conditions remains a great challenge due to the presence of complex matrix and extremely small cell volumes. Herein, a serpentine channel microfluidic device which was designed to achieve continuous cell separation and inertial focusing, was coupled with a pulsed electric field-induced electrospray ionization-high resolution MS (PEF-ESI-HRMS) to achieve high-throughput single cell analysis. The pulsed square wave electric field was applied to realize on-line cell disruption and induce electrospray ionization. Single cells were analyzed under near-physiological conditions at a throughput of up to 80 cells min^-1. More than 900 features were detected and approximately 120 metabolites were tentatively identified from a single cell. Further, by continually analyzing more than 3000 MCF7 and HepG2 cells, discrimination of different cancer cells based on their individual metabolic profiles was achieved by using the principal component analysis. The PEF-ESI-HRMS method was also applied for the analysis of single yeast cells, and more than 40 metabolites were annotated. This method is versatile and has good robustness, which is promising for high-throughput single cell metabolomics analysis.

Keywords: Cellular heterogeneity; High-throughput; Mass spectrometry; Microfluidic device; Single cell metabolomics.

MeSH terms

Cell Separation
Metabolome
Metabolomics / methods
Microfluidics*
Spectrometry, Mass, Electrospray Ionization* / methods