Single-Cell Mass Spectrometry of Metabolites Extracted from Live Cells by Fluidic Force Microscopy

Orane Guillaume-Gentil, Timo Rey, Patrick Kiefer, Alfredo J. Ibáñez, Robert Steinhoff, Rolf Brönnimann, Livie Dorwling-Carter, Tomaso Zambelli, Renato Zenobi, Julia A. Vorholt

Producción científica: Contribución a una revistaArtículorevisión exhaustiva

80 Citas (Scopus)

Resumen

Single-cell metabolite analysis provides valuable information on cellular function and response to external stimuli. While recent advances in mass spectrometry reached the sensitivity required to investigate metabolites in single cells, current methods commonly isolate and sacrifice cells, inflicting a perturbed state and preventing complementary analyses. Here, we propose a two-step approach that combines nondestructive and quantitative withdrawal of intracellular fluid with subpicoliter resolution using fluidic force microscopy, followed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The developed method enabled the detection and identification of 20 metabolites recovered from the cytoplasm of individual HeLa cells. The approach was further validated in 13C-glucose feeding experiments, which showed incorporation of labeled carbon atoms into different metabolites. Metabolite sampling, followed by mass spectrometry measurements, enabled the preservation of the physiological context and the viability of the analyzed cell, providing opportunities for complementary analyses of the cell before, during, and after metabolite analysis.

Idioma originalInglés
Páginas (desde-hasta)5017-5023
Número de páginas7
PublicaciónAnalytical Chemistry
Volumen89
N.º9
DOI
EstadoPublicada - 2 may. 2017
Publicado de forma externa

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