Single and few cell analysis for correlative light microscopy, metabolomics, and targeted proteomics
Luca Rima, Christian Berchtold, Stefan Arnold, Andri Fränkl, Rosmarie Sütterlin, Gregor Dernick, Götz Schlotterbeck, Thomas Braun
TL;DR
This paper introduces a method to study single or few cells by combining microscopy with metabolomics and proteomics to understand cellular processes.
Contribution
A new method for correlative single-cell analysis combining light microscopy, metabolomics, and proteomics.
Findings
The method enables correlative measurement of cellular structures and metabolites.
It allows for targeted protein detection on the single-cell level using RPPA.
The approach combines LC-MS and RPPA for few-cell analysis.
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…
Genes, proteins, chemicals, diseases, species, mutations and cell lines named across the full text — each resolved to its canonical identifier and authoritative record.
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Taxonomy
TopicsSingle-cell and spatial transcriptomics · Cell Image Analysis Techniques · Advanced Biosensing Techniques and Applications