期刊
ANALYTICAL CHEMISTRY
卷 89, 期 9, 页码 5017-5023出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.7b00367
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资金
- ETH Zurich
- Swiss Innovation Promotion Agency CTI-KTI [14336.1 PFNM-NM, 18511.1 PFNM-NM]
- Swiss National Science Foundation [PZ00P3_142615]
- Swiss National Science Foundation (SNF) [PZ00P3_142615] Funding Source: Swiss National Science Foundation (SNF)
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 C-13-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
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