Determination of intracellular species at the level of a single erythrocyte via capillary electrophoresis with direct and indirect fluorescence detection

Hogan, Barry L.; Yeung, Edward S.

doi:10.1021/ac00046a031

Cited by 221 publications

(150 citation statements)

References 22 publications

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“…Capillary electrophoresis is an emerging technique capable of rapidly determining multiple chemical species in picoliter and femtoliter biological samples, including single cells (5)(6)(7)(8)(9)(10). This technique has been used successfully to develop quantitative analyses for chemicals in single whole nerve cells, including large invertebrate cells (8) and small mammalian erythrocytes (9).…”

Section: Activitymentioning

confidence: 99%

“…This technique has been used successfully to develop quantitative analyses for chemicals in single whole nerve cells, including large invertebrate cells (8) and small mammalian erythrocytes (9). Capillary electrophoresis has been used to remove picoliter samples of cytoplasm from single large invertebrate neurons for separation and quantitative detection by capillary electrophoresis (10).…”

Section: Activitymentioning

confidence: 99%

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Discovery of endogenous catecholamines in lymphocytes and evidence for catecholamine regulation of lymphocyte function via an autocrine loop.

Bergquist¹,

Tarkowski²,

Ekman³

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

345

262

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Evidence has been obtained that catecholamines and their metabolites are present in single lymphocytes and extracts of T-and B-ceUl clones by use of capiUlary electrophoresis with electrochemical detection. Pharmacological inhibition of tyrosine hydroxylase reduces observed catecholamine levels, suggesting catecholamine synthesis by lymphocytes. Intracelular dopamine levels are shown to be increased by extracellular dopamine, suggesting a cellular-uptake mechanism. Furthermore, incubation with either dopamine or L-dihydroxyphenylalanine, a precursor of dopamine, results in a dosedependent inhibition of lymphocyte proliferation and differentiation. Together, these results suggest the presence of an autocrine loop whereby lymphocytes down-regulate their own activity.

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Section: Activitymentioning

confidence: 99%

Section: Activitymentioning

confidence: 99%

Discovery of endogenous catecholamines in lymphocytes and evidence for catecholamine regulation of lymphocyte function via an autocrine loop.

Bergquist¹,

Tarkowski²,

Ekman³

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

345

262

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show abstract

“…Researchers must investigate the reactivity of a protein with its physiological partners to understand its role in cellular pathways; concentration measurements alone could be misleading, because the local environment of a protein can drastically alter its activity. Emerging capillary micro-/nanofabrication technologies allow the chemical reactivity of the target species to be investigated (3,(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21). Allbritton and coworkers (11,12) embarked on the development of this technology and used capillary electrophoresis to analyze intracellular kinase activity.…”

mentioning

confidence: 99%

Nanokit for single-cell electrochemical analyses

Pan

Jiang

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

115

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The development of more intricate devices for the analysis of small molecules and protein activity in single cells would advance our knowledge of cellular heterogeneity and signaling cascades. Therefore, in this study, a nanokit was produced by filling a nanometersized capillary with a ring electrode at the tip with components from traditional kits, which could be egressed outside the capillary by electrochemical pumping. At the tip, femtoliter amounts of the kit components were reacted with the analyte to generate hydrogen peroxide for the electrochemical measurement by the ring electrode. Taking advantage of the nanotip and small volume injection, the nanokit was easily inserted into a single cell to determine the intracellular glucose levels and sphingomyelinase (SMase) activity, which had rarely been achieved. High cellular heterogeneities of these two molecules were observed, showing the significance of the nanokit. Compared with the current methods that use a complicated structural design or surface functionalization for the recognition of the analytes, the nanokit has adapted features of the well-established kits and integrated the kit components and detector in one nanometer-sized capillary, which provides a specific device to characterize the reactivity and concentrations of cellular compounds in single cells.

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“…ATP is known to evoke intercellular calcium wave propagation from glial [26,112] and other [50] cells. The astrocyte cultures were loaded with fluo-3 [60], and the fluorescence on binding with intracellular Ca, [Ca 2+ ], was monitored simultaneously with the chemiluminescence signal from the released ATP.…”

Section: Atp In Central Nervous Systemmentioning

confidence: 99%

Selective detection of neurotransmitters by fluorescence and chemiluminescence imaging

Wang

Yeung

2001

Pure and Applied Chemistry

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In recent years, luminescence imaging has been widely employed in neurochemical analysis. It has a number of advantages for the study of neuronal and other biological cells: (1) a particular molecular species or cellular constituent can be selectively visualized in the presence of a large excess of other species in a heterogeneous environment; (2) low concentration detection limits can be achieved because of the inherent sensitivity associated with fluorescence and chemiluminescence; (3) low excitation intensities can be used so that long-term observation can be realized while the viability of the specimen is preserved; and (4) excellent spatial resolution can be obtained with the light microscope so subcellular compartments can be identified. With good sensitivity, temporal and spatial resolution, the flux of ions and molecules and the distribution and dynamics of intracellular species can be measured in real time with specific luminescence probes, substrates, or with native fluorescence. A noninvasive detection scheme based on glutamate dehydrogenase (GDH) enzymatic assay combined with microscopy was developed to measure the glutamate release in cultured cells from the central nervous system (CNS). The enzyme reaction is very specific and sensitive. The detection limit with CCD imaging is down to µM levels of glutamate with reasonable response time. We also found that chemiluminescence associated with the ATP-dependent reaction between luciferase and luciferin can be used to image ATP at levels down to 10 nM in the millisecond time scale. Similar imaging experiments should be feasible in a broad spectrum of biological systems.

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Determination of intracellular species at the level of a single erythrocyte via capillary electrophoresis with direct and indirect fluorescence detection

Cited by 221 publications

References 22 publications

Discovery of endogenous catecholamines in lymphocytes and evidence for catecholamine regulation of lymphocyte function via an autocrine loop.

Discovery of endogenous catecholamines in lymphocytes and evidence for catecholamine regulation of lymphocyte function via an autocrine loop.

Nanokit for single-cell electrochemical analyses

Selective detection of neurotransmitters by fluorescence and chemiluminescence imaging

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