Single-cell mass spectrometry imaging of lipids in HeLa cells via tapping-mode scanning probe electrospray ionization

Abstract The development of analytical technology that allows investigation of the diversity of cells that form biological tissues based on molecular information is important to elucidate the heterogeneity of cells and pathological mechanisms. Here, we present a proof-of-concept demonstration of sin...

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Main Authors: Yoichi Otsuka, Kazuya Kabayama, Ayane Miura, Masatomo Takahashi, Kosuke Hata, Yoshihiro Izumi, Takeshi Bamba, Koichi Fukase, Michisato Toyoda
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:Communications Chemistry
Online Access:https://doi.org/10.1038/s42004-025-01521-2
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author Yoichi Otsuka
Kazuya Kabayama
Ayane Miura
Masatomo Takahashi
Kosuke Hata
Yoshihiro Izumi
Takeshi Bamba
Koichi Fukase
Michisato Toyoda
author_facet Yoichi Otsuka
Kazuya Kabayama
Ayane Miura
Masatomo Takahashi
Kosuke Hata
Yoshihiro Izumi
Takeshi Bamba
Koichi Fukase
Michisato Toyoda
author_sort Yoichi Otsuka
collection DOAJ
description Abstract The development of analytical technology that allows investigation of the diversity of cells that form biological tissues based on molecular information is important to elucidate the heterogeneity of cells and pathological mechanisms. Here, we present a proof-of-concept demonstration of single-cell mass spectrometry imaging (SC-MSI) via tapping-mode scanning probe electrospray ionization (t-SPESI), which is an atmospheric-pressure sampling ionization technique. We developed a novel t-SPESI unit that can be used in combination with an inverted fluorescence microscope and basic technologies to extract components from microregions of cells and measure ions with high sensitivity. We performed multimodal (fluorescence, lipid ion, and topographic) imaging of two types of HeLa cells labeled with fluorescent dyes and chemically fixed and showed the potential for subcellular-scale analysis of both cell structure and chemical composition. Furthermore, we evaluated the lipid species by comparing the SC-MSI results with those of supercritical fluid chromatography tandem mass spectrometry. The technical advancement presented here is effective for distinguishing cell types based on the signal intensity of lipid ions in single cells and investigating differences in the subcellular localization of lipids in different types of cells.
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spelling doaj-art-e32427a142b54288ab6642e4e07bb91c2025-08-20T02:34:06ZengNature PortfolioCommunications Chemistry2399-36692025-05-018111210.1038/s42004-025-01521-2Single-cell mass spectrometry imaging of lipids in HeLa cells via tapping-mode scanning probe electrospray ionizationYoichi Otsuka0Kazuya Kabayama1Ayane Miura2Masatomo Takahashi3Kosuke Hata4Yoshihiro Izumi5Takeshi Bamba6Koichi Fukase7Michisato Toyoda8Department of Physics, Graduate School of Science, The University of OsakaDepartment of Chemistry, Graduate School of Science, The University of OsakaDepartment of Chemistry, Graduate School of Science, The University of OsakaDivision of Metabolomics, Medical Research Center for High Depth Omics, Medical Institute of Bioregulation, Kyushu UniversityDivision of Metabolomics, Medical Research Center for High Depth Omics, Medical Institute of Bioregulation, Kyushu UniversityDivision of Metabolomics, Medical Research Center for High Depth Omics, Medical Institute of Bioregulation, Kyushu UniversityDivision of Metabolomics, Medical Research Center for High Depth Omics, Medical Institute of Bioregulation, Kyushu UniversityDepartment of Chemistry, Graduate School of Science, The University of OsakaDepartment of Physics, Graduate School of Science, The University of OsakaAbstract The development of analytical technology that allows investigation of the diversity of cells that form biological tissues based on molecular information is important to elucidate the heterogeneity of cells and pathological mechanisms. Here, we present a proof-of-concept demonstration of single-cell mass spectrometry imaging (SC-MSI) via tapping-mode scanning probe electrospray ionization (t-SPESI), which is an atmospheric-pressure sampling ionization technique. We developed a novel t-SPESI unit that can be used in combination with an inverted fluorescence microscope and basic technologies to extract components from microregions of cells and measure ions with high sensitivity. We performed multimodal (fluorescence, lipid ion, and topographic) imaging of two types of HeLa cells labeled with fluorescent dyes and chemically fixed and showed the potential for subcellular-scale analysis of both cell structure and chemical composition. Furthermore, we evaluated the lipid species by comparing the SC-MSI results with those of supercritical fluid chromatography tandem mass spectrometry. The technical advancement presented here is effective for distinguishing cell types based on the signal intensity of lipid ions in single cells and investigating differences in the subcellular localization of lipids in different types of cells.https://doi.org/10.1038/s42004-025-01521-2
spellingShingle Yoichi Otsuka
Kazuya Kabayama
Ayane Miura
Masatomo Takahashi
Kosuke Hata
Yoshihiro Izumi
Takeshi Bamba
Koichi Fukase
Michisato Toyoda
Single-cell mass spectrometry imaging of lipids in HeLa cells via tapping-mode scanning probe electrospray ionization
Communications Chemistry
title Single-cell mass spectrometry imaging of lipids in HeLa cells via tapping-mode scanning probe electrospray ionization
title_full Single-cell mass spectrometry imaging of lipids in HeLa cells via tapping-mode scanning probe electrospray ionization
title_fullStr Single-cell mass spectrometry imaging of lipids in HeLa cells via tapping-mode scanning probe electrospray ionization
title_full_unstemmed Single-cell mass spectrometry imaging of lipids in HeLa cells via tapping-mode scanning probe electrospray ionization
title_short Single-cell mass spectrometry imaging of lipids in HeLa cells via tapping-mode scanning probe electrospray ionization
title_sort single cell mass spectrometry imaging of lipids in hela cells via tapping mode scanning probe electrospray ionization
url https://doi.org/10.1038/s42004-025-01521-2
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