Single and few cell analysis for correlative light microscopy, metabolomics, and targeted proteomics
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Author (Corporation)
Publication date
08.2024
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Type
01A - Journal article
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Editor (Corporation)
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Parent work
Lab on a Chip
Special issue
DOI of the original publication
Link
Series
Series number
Volume
24
Issue / Number
18
Pages / Duration
4321-4332
Patent number
Publisher / Publishing institution
Royal Society of Chemistry
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Abstract
The interactions of proteins, membranes, nucleic acid, and metabolites shape a cell's phenotype. These interactions are stochastic, and each cell develops differently, making it difficult to synchronize cell populations. Consequently, studying biological processes at the single- or few-cell level is often necessary to avoid signal dilution below the detection limit or averaging over many cells. We have developed a method to study metabolites and proteins from a small number of or even a single adherent eukaryotic cell. Initially, cells are lysed by short electroporation and aspirated with a microcapillary under a fluorescent microscope. The lysate is placed on a carrier slide for further analysis using liquid-chromatography mass spectrometry (LC-MS) and/or reverse-phase protein (RPPA) approach. This method allows for a correlative measurement of (i) cellular structures and metabolites and (ii) cellular structures and proteins on the single-cell level. The correlative measurement of cellular structure by light-microscopy, metabolites by LC-MS, and targeted protein detection by RPPA was possible on the few-cell level. We discuss the method, potential applications, limitations, and future improvements.
Keywords
Animals, Liquid chromatography, Humans, Mass spectrometry, Metabolomics, Microscopy, Proteomics, Single-cell analysis
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ISBN
ISSN
1473-0197
1473-0189
1473-0189
Language
English
Created during FHNW affiliation
Yes
Strategic action fields FHNW
Publication status
Published
Review
Peer review of the complete publication
Open access category
Hybrid
License
Citation
Rima, L., Berchtold, C., Arnold, S., Fränkl, A., Sütterlin, R., Dernick, G., Schlotterbeck, G., & Braun, T. (2024). Single and few cell analysis for correlative light microscopy, metabolomics, and targeted proteomics. Lab on a Chip, 24(18), 4321–4332. https://doi.org/10.1039/d4lc00269e