Issue 18, 2024

Single and few cell analysis for correlative light microscopy, metabolomics, and targeted proteomics

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.

Graphical abstract: Single and few cell analysis for correlative light microscopy, metabolomics, and targeted proteomics

Supplementary files

Article information

Article type
Paper
Submitted
26 Mar 2024
Accepted
23 Jul 2024
First published
08 Aug 2024
This article is Open Access
Creative Commons BY license

Lab Chip, 2024,24, 4321-4332

Single and few cell analysis for correlative light microscopy, metabolomics, and targeted proteomics

L. Rima, C. Berchtold, S. Arnold, A. Fränkl, R. Sütterlin, G. Dernick, G. Schlotterbeck and T. Braun, Lab Chip, 2024, 24, 4321 DOI: 10.1039/D4LC00269E

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