Determination of glucose in human stomach cancer cell extracts and single cells by capillary electrophoresis with a micro-biosensor

Wang, Xiaolei; Ma, Yu; Zhao, Man; Zhou, Meiling; Yan, Xiao; Sun, Zifei; Tong, Lili

doi:10.1016/j.chroma.2016.09.054

Cited by 29 publications

(13 citation statements)

References 37 publications

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“…[34] CE-LIF and CE-ESI-MS of individual cells remain active areas of research, with samples ranging from developing Xenopus laevis embryos [35] to human stomach cancer cells. [36] Recently,t he Nemes group [37] quantified proteins using bottom-up CE-ESI-MS and detected over 438 non-redundant protein groups during different developmental stages of the X. laevis embryo. Further,t hey studied the metabolic contents within certain blastomeres that contribute to cell differentiation into neuronal, epidermal, and hindgut tissues within 16-cell X. laevis embryos.O verall, 80 metabolites were detected and used to develop predictive metabolic profiles for undifferentiated cells.T his same group further investigated temporal changes by utilizing live-cell CE-MS to probe metabolic differences between the dorsal and ventral side of X. laevis embryos at eight-and sixteen-cell stages.M etabolic differentiation oc-Angewandte Chemie Reviews 9352 www.angewandte.org curred as early as eight cells,w hich is earlier than originally predicted.…”

Section: Comprehensive Multimodal Chemical Analysismentioning

confidence: 99%

Exploring the Fundamental Structures of Life: Non‐Targeted, Chemical Analysis of Single Cells and Subcellular Structures

et al. 2019

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Cells are a basic functional and structural unit of living organisms. Both unicellular communities and multicellular species produce an astonishing chemical diversity, enabling a wide range of divergent functions, yet each cell shares numerous aspects that are common to all living organisms. While there are many approaches for studying this chemical diversity, only a few are non‐targeted and capable of analyzing hundreds of different chemicals at cellular resolution. Here, we review the non‐targeted approaches used to perform comprehensive chemical analyses, provide chemical imaging information, or obtain high‐throughput single‐cell profiling data. Single‐cell measurement capabilities are rapidly increasing in terms of throughput, limits of detection, and completeness of the chemical analyses; these improvements enable their application to understand ever more complex physiological phenomena, such as learning, memory, and behavior.

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Section: Comprehensive Multimodal Chemical Analysismentioning

confidence: 99%

Exploring the Fundamental Structures of Life: Non‐Targeted, Chemical Analysis of Single Cells and Subcellular Structures

et al. 2019

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“…[24] Die mechanische oder enzymatische Entfernung von Zellen kann zu Verformungen und der Freisetzung wichtiger Verbindungen führen. [34] CE-LIF und CE-ESI-MS einzelner Zellen bleiben aktive Forschungsgebiete,w obei die Proben von sich entwickelnden Embryonen innerhalb der Spezies Xenopus laevis [35] bis zu menschlichen Magenkrebszellen [36] reichen. Ein alternatives,s chnelles Lyseschema verwendet eine Elektrode,umeine Potentialdifferenz von 30 V über die Zellmembran zu induzieren.…”

Section: Umfassende Multimodale Chemische Analyseunclassified

“…[29] [30] Die nicht zielgerichtete CE-MS-Analyse wurde erfolgreich eingesetzt, um den Stoffwechselgehalt in einer Vielfalt biologischer Systeme,wie Neuronen, [31] Krebszellen, [32] Pflanzen [33] und roten Blutkçrperchen, zu bestimmen. [34] CE-LIF und CE-ESI-MS einzelner Zellen bleiben aktive Forschungsgebiete,w obei die Proben von sich entwickelnden Embryonen innerhalb der Spezies Xenopus laevis [35] bis zu menschlichen Magenkrebszellen [36] reichen. Vor kurzem quantifizierten Nemes und Mitarbeiter [37] Proteine unter Verwendung von "bottom up"-CE-ESI-MS,wobei über 438 nichtredundante Proteingruppen in verschiedenen Entwicklungsstadien des X. laevis-Embryos nachgewiesen wurden.…”

Section: Umfassende Multimodale Chemische Analyseunclassified

Erforschung der fundamentalen Strukturen des Lebens: Nicht zielgerichtete chemische Analyse von Einzelzellen und subzellulären Strukturen

et al. 2019

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Zellen sind die grundlegende Funktions‐ und Struktureinheit lebender Organismen. Einzellige Gemeinschaften wie auch mehrzellige Spezies produzieren eine erstaunliche chemische Diversität, die eine große Bandbreite an unterschiedlichen Funktionen ermöglicht, dennoch hat jede Zelle zahlreiche Merkmale, die allen lebenden Organismen gemeinsam sind. Während es viele Ansätze zur Erforschung dieser chemischen Diversität gibt, sind nur wenige nicht zielgerichtet und in der Lage, die Analyse von Hunderten von verschiedenen Substanzen mit zellulärer Auflösung zu bewerkstelligen. In diesem Aufsatz diskutieren wir solche nicht zielgerichteten Ansätze, die verwendet werden, um umfassende chemische Analysen durchzuführen, Informationen für die chemische Bildgebung bereitzustellen oder Hochdurchsatz‐Profilingdaten über einzelne Zellen zu erhalten. Die Fähigkeiten der Einzelzellmessung entwickeln sich rasant bezüglich Durchsatz, Nachweisgrenzen und der Vollständigkeit der chemischen Analysen; diese Verbesserungen ermöglichen es, immer komplexere physiologische Phänomene, wie z. B. Lernen, Gedächtnis und Verhalten zu verstehen.

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“…The benefits of single-cell analysis have become increasingly apparent in recent years as heterogeneity among cells in a population has been identified [13–15]. A variety of cellular metabolites have been assayed using single-cell CZE, including thiol-containing compounds [16], small molecules such as glucose [17], dopamine [18], and amino acids [19], as well as measures of enzyme and peptidase activity [2023]. Single-cell CZE has also been used to study different classes of lipid metabolism in cytosolic lysates and single cells [24], such as sphingolipids [25,26], glycosphingolipids [27], and phosphatidylinositides [28].…”

Section: Introductionmentioning

confidence: 99%

Chemical fixation to arrest phospholipid signaling for chemical cytometry

Proctor

Sims

Allbritton

2017

Journal of Chromatography A

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Chemical cytometry is a powerful tool for measuring biological processes such as enzymatic signaling at the single cell level. Among these technologies, single-cell capillary zone electrophoresis (CZE) has emerged as a powerful tool to assay a wide range of cellular metabolites. However, analysis of dynamic processes within cells remains challenging as signaling pathways are rapidly altered in response to changes in the cellular environment, including cell manipulation and storage. To address these limitations, we describe a method for chemical fixation of cells to stop the cellular reactions to preserve the integrity of key signaling molecules or reporters within the cell and to enable the cell to act as a storage reservoir for the reporter and its metabolites prior to assay by single-cell CZE. Fluorescent phosphatidylinositol 4,5-bisphosphate reporters were loaded into cells and the cells were chemically fixed and stored prior to analysis. The reporter and its metabolites were electrophoretically separated by singlecell CZE. Chemical fixation parameters such as fixative, fixation time, storage solution, storage duration, and extraction solution were optimized. When cells were loaded with a fluorescent C6- or C16-PIP2 followed by glutaraldehyde fixation and immediate analysis, 24 ± 2% and 139 ± 12% of the lipid was recoverable, respectively, when compared to an unfixed control. Storage of the cells for 24 h yielded recoverable lipid of 61 ± 3% (C6-PIP2) and 55 ± 5% (C16-PIP2) when compared to cells analyzed immediately after fixation. The metabolites observed with and without fixation were identical. Measurement of phospholipase C activity in single leukemic cells in response to an agonist demonstrated the capability of chemical fixation coupled to singlecell CZE to yield an accurate snapshot of cellular reactions with the probe. This methodology enables cell assay with the reporter to be separated in space and time from reporter metabolite quantification while preserving assay integrity.

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Determination of glucose in human stomach cancer cell extracts and single cells by capillary electrophoresis with a micro-biosensor

Cited by 29 publications

References 37 publications

Exploring the Fundamental Structures of Life: Non‐Targeted, Chemical Analysis of Single Cells and Subcellular Structures

Exploring the Fundamental Structures of Life: Non‐Targeted, Chemical Analysis of Single Cells and Subcellular Structures

Erforschung der fundamentalen Strukturen des Lebens: Nicht zielgerichtete chemische Analyse von Einzelzellen und subzellulären Strukturen

Chemical fixation to arrest phospholipid signaling for chemical cytometry

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