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Global and Spatial Metabolomics of Individual Cells Using a Tapered Pneumatically Assisted nano-DESI Probe
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - BMC, Analytical Chemistry.ORCID iD: 0000-0002-8433-1725
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - BMC, Analytical Chemistry.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Cell Biology.ORCID iD: 0000-0003-2918-8142
Karolinska Inst, Dept Biosci & Nutr, S-14152 Huddinge, Sweden..
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2023 (English)In: Journal of the American Society for Mass Spectrometry, ISSN 1044-0305, E-ISSN 1879-1123, Vol. 34, no 11, p. 2518-2524Article in journal (Refereed) Published
Abstract [en]

Single-cell metabolomics has the potential to reveal unique insights into intracellular mechanisms and biological processes. However, the detection of metabolites from individual cells is challenging due to their versatile chemical properties and concentrations. Here, we demonstrate a tapered probe for pneumatically assisted nanospray desorption electrospray ionization (PA nano-DESI) mass spectrometry that enables both chemical imaging of larger cells and global metabolomics of smaller 15 mu m cells. Additionally, by depositing cells in predefined arrays, we show successful metabolomics from three individual INS-1 cells per minute, which enabled the acquisition of data from 479 individual cells. Several cells were used to optimize analytical conditions, and 93 or 97 cells were used to monitor metabolome alterations in INS-1 cells after exposure to a low or high glucose concentration, respectively. Our analytical approach offers insights into cellular heterogeneity and provides valuable information about cellular processes and responses in individual cells.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2023. Vol. 34, no 11, p. 2518-2524
National Category
Analytical Chemistry Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-516884DOI: 10.1021/jasms.3c00239ISI: 001097001800001PubMedID: 37830184OAI: oai:DiVA.org:uu-516884DiVA, id: diva2:1817808
Funder
Swedish Research Council, 2017-04125EU, European Research Council, 101041224 - X CELLSwedish Research Council, VR MH 2019-02050Novo Nordisk, NNF21OC0070086Novo Nordisk, NNF22OC0078239Swedish Society for Medical Research (SSMF)Available from: 2023-12-07 Created: 2023-12-07 Last updated: 2024-04-24Bibliographically approved
In thesis
1. Down to single-cell metabolomics: Developments of direct infusion tools for electrospray ionization mass spectrometry
Open this publication in new window or tab >>Down to single-cell metabolomics: Developments of direct infusion tools for electrospray ionization mass spectrometry
2024 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Metabolomics provides a snapshot of the metabolic status of biological samples. This knowledge can give insights into several biological processes, including understanding the etiology and development of diseases such as cancer or type 2 diabetes. Mass spectrometry is the golden standard technique for metabolite profiling due to its high sensitivity and comprehensive analyte coverage. Although several sampling techniques are available for routine metabolomics studies, these usually require sample sizes of around 1 million cells. The work entailed in this thesis focuses on developing sampling strategies to perform metabolomic studies on challenging samples, namely, low cell number and minute volume samples, down to single cells. 

The direct infusion probe (DIP) that I developed enables the analysis of metabolites and lipids in minute sample volumes with low cell numbers. Specifically, it enables analysis of samples containing as low as 20 cells/µL in 5 µL volumes. The DIP has a user-friendly and simple design that allows for rapidly switching samples. Furthermore, it provides minimal carryover between samples by simply washing with the next sample for 8 seconds, which enhances the analysis throughput by eliminating extra washing steps. I utilized the DIP to conduct metabolite and lipid profiling of insulin-releasing cells (INS-1) exposed to high glucose. I observed that high glucose exposure causes changes in the abundance of amino acids, lipids, and other small molecules. Our results are linked with pathways, such as glycolysis, fatty acid biosynthesis, and glutamate and sphingolipid metabolisms that are altered in type 2 diabetes. 

Another analytically challenging task that I address in this thesis is the chemical analysis of individual cells. Single-cell metabolomics is crucial to decoding cellular heterogeneity and identifying phenotype variations within a cell population. I redesigned the pneumatically assisted nanospray desorption electrospray ionization (PA nano-DESI) probe to enable the analysis of metabolites and lipids from single cells. This was accomplished by reducing the inner diameter of capillaries to enhance ionization and tapering the capillaries of the probe to establish a miniaturized liquid bridge. Then, I utilized the tapered PA nano-DESI probe to profile the metabolome and lipidome of single INS-1 cells and senescent IMR-90 cells. The results show alterations in glutamate and arachidonic acid levels, characteristic of insulin release and aging processes, respectively. 

Overall, the DIP and the tapered PA nano-DESI probes represent significant analytical advances in conducting metabolomic studies of samples containing low cell numbers down to single cells.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2024. p. 85
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 2406
Keywords
mass spectrometry, single-cell, metabolomics, lipidomics, DIP-MS, tapered PA-nano-DESI
National Category
Analytical Chemistry
Research subject
Chemistry with specialization in Analytical Chemistry
Identifiers
urn:nbn:se:uu:diva-527116 (URN)978-91-513-2142-4 (ISBN)
Public defence
2024-06-13, A1:107a, BMC, Husargatan 3, Uppsala, 09:15 (English)
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Supervisors
Available from: 2024-05-22 Created: 2024-04-24 Last updated: 2024-05-22

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Marques-Santos, Cátia M.Friedrich, FelixLiu, LiangwenLanekoff, Ingela

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