Chemical Analysis of Single Cells

Lin, Yuqing; Trouillon, Raphaël; Safina, Gulnara; Ewing, Andrew G.

doi:10.1021/ac2009838

Cited by 178 publications

(137 citation statements)

References 292 publications

(544 reference statements)

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“…The measurements were performed in Hepes-based saline solution (10 mM Hepes, 150 mM NaCl, 4.2 mM KCl, and 11.2 mM glucose; pH 7.4). In this experiment, we used relatively large electrodes (a ¼ 6.0 μm) to detect neurotransmitter effectively, based on the rationale outlined in earlier work for both NO and neurotransmitter detection (43)(44)(45)(46)(47)(48). The carbon electrode was simply aligned optically over the cell and positioned one radius above the cell surface using the reduction current at a potential of −1.0 V, due to oxygen reduction, which was found to provide reliable distance control.…”

Section: Resultsmentioning

confidence: 99%

Topographical and electrochemical nanoscale imaging of living cells using voltage-switching mode scanning electrochemical microscopy

Takahashi

Shevchuk

Novák

et al. 2012

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

206

224

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We describe voltage-switching mode scanning electrochemical microscopy (VSM-SECM), in which a single SECM tip electrode was used to acquire high-quality topographical and electrochemical images of living cells simultaneously. This was achieved by switching the applied voltage so as to change the faradaic current from a hindered diffusion feedback signal (for distance control and topographical imaging) to the electrochemical flux measurement of interest. This imaging method is robust, and a single nanoscale SECM electrode, which is simple to produce, is used for both topography and activity measurements. In order to minimize the delay at voltage switching, we used pyrolytic carbon nanoelectrodes with 6.5-100 nm radii that rapidly reached a steady-state current, typically in less than 20 ms for the largest electrodes and faster for smaller electrodes. In addition, these carbon nanoelectrodes are suitable for convoluted cell topography imaging because the RG value (ratio of overall probe diameter to active electrode diameter) is typically in the range of 1.5-3.0. We first evaluated the resolution of constant-current mode topography imaging using carbon nanoelectrodes. Next, we performed VSM-SECM measurements to visualize membrane proteins on A431 cells and to detect neurotransmitters from a PC12 cells. We also combined VSM-SECM with surface confocal microscopy to allow simultaneous fluorescence and topographical imaging. VSM-SECM opens up new opportunities in nanoscale chemical mapping at interfaces, and should find wide application in the physical and biological sciences.high-resolution imaging | living cell imaging | noninvasive | constant-distance mode S canning electrochemical microscopy (SECM) uses an electrode tip for detecting electroactive chemical species and is an effective tool for the investigation of the localized chemical properties of sample surfaces and interfaces (1). Because SECM has high temporal resolution and can be used under physiological conditions, it is particularly well suited for quantitative measurements of (short-lived) chemicals like neurotransmitters, nitric oxide, reactive oxygen species, and oxygen, which are released/ consumed by living cells. In conventional SECM, the probe is often micrometer scale and the probe vertical position is kept at a constant height, a plane, during probe scanning. If the sample topography is not flat, the electrode-sample separation changes during scanning, complicating the SECM measurement and its analysis.Various methods for SECM electrode miniaturization and control of the electrode-sample separation have been advocated in order to improve the resolution of SECM imaging. The reliable fabrication of nanoelectrodes with a small ratio of electrodeinsulation to active electrode (

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Section: Resultsmentioning

confidence: 99%

Topographical and electrochemical nanoscale imaging of living cells using voltage-switching mode scanning electrochemical microscopy

Takahashi

Shevchuk

Novák

et al. 2012

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

206

224

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“…3 In addition, the concentration of these trace elements can have a large effect on their toxicity within biological systems, making accurate elemental analysis of cells an essential technique. 4 The field of research regarding analysis of metal-containing biomolecules, "metallomes", was termed "Metallomics" by Haraguchi in 2004, 1 and a respective journal also came into publication in 2009. 5 In the relevant review he stated that the "all-elements analysis of one biological cell is now a challenging subject of analytical biochemistry", and went on to describe the multielement analysis of salmon egg cells with inductively coupled plasma optical emission spectrometry (ICP-OES) and inductively coupled plasma mass spectrometry (ICP-MS) after acid digestion, as an example of research into this field.…”

Section: Introductionmentioning

confidence: 99%

“…In recent years, single-cell elemental analysis using ICP-OES and ICP-MS has gained substantial interest in the biochemical and metallomics fields. 1,4,[7][8][9][10][11][12] If a suspension of cells can be directly introduced into the plasma as an aerosol, detailed information regarding the distribution of elements within a sample of cells can be obtained. There are many possible applications of this, such as simply comparing phosphorus levels in biological cells to give the distribution of cell size in a sample.…”

Section: Introductionmentioning

confidence: 99%

High Sensitive Elemental Analysis of Single Yeast Cells (Saccharomyces cerevisiae) by Time-Resolved Inductively-Coupled Plasma Mass Spectrometry Using a High Efficiency Cell Introduction System

et al. 2013

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Trace elemental analysis of single yeast cells with time-resolved inductively coupled plasma mass spectrometry (ICP-MS) was successfully carried out, where a high efficiency cell introduction system (HECIS) consisting of the high performance concentric nebulizer (HPCN) and a low-volume (15 mL) on-axis spray chamber utilizing a sheath gas flow were used. Cell adsorption to the flow injector and sample tubing was reduced with the addition of a simple 4.3 mmol L -1 of NaCl solution to the cell suspension and cell flowing liquid, allowing consecutive measurements without fear of significant contamination from previous measurements. Initially using a quadrupole mass analyzer ICP-MS (ICP-QMS) at its lowest integration time (10 ms), current spikes corresponding to separate cell events were detected for several elements (Mg, P, Ca, Mn, Fe, Cu, and Zn) on the introduction of the cell suspension. On comparing the number of peaks in the spectrum for phosphorous with the cell count using a haemocytometer, a reproducible cell transport efficiency of 75.0 ± 4.7% was achieved. Preliminary experiments into using time of flight ICP-MS (ICP-TOFMS) for single-cell analysis were carried out, allowing quasi-simultaneous multielement detection. The spectra of Mg, P, Ca, Mn, Fe, Cu, and Zn, with a time resolution of 1 ms were simultaneously obtained in one measurement. A relatively strong correlation was observed for the spectra between P and Zn (correlation factor 0.69), P and Mg (0.63), and Mg and Zn (0.63). These results indicate that the time resolved quasi-simultaneous multielement measurement may be useful for the correlation analysis of multielements in cells.

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“…Due to cellular heterogeneity within an isogenic cell population, individual analysis of cells will lead to a more sensitive representation of cell-to-cell variations, instead of a stochastic average analysis by bulk measurements. [24][25][26][27][28][29] Classical ICP-MS analysis of cells is the bulk analysis of large amounts of cells with a lysis, extraction or digestion; 30 this is an approach that is hardly adapted for evaluating any cell-to-cell variance. Time-resolved measurement via a direct cell introduction method is a relatively recent method for individual analysis of cells by ICP-MS, [14][15][16][17][18][19][20][21][22][23] an approach that can detect the elements in individual cells with a high sensitivity, often called "single particle mode".…”

Section: Special Reviewsmentioning

confidence: 99%

Time-resolved ICP-MS Measurement: a New Method for Elemental and Multiparametric Analysis of Single Cells

et al. 2014

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Chemical Analysis of Single Cells

Cited by 178 publications

References 292 publications

Topographical and electrochemical nanoscale imaging of living cells using voltage-switching mode scanning electrochemical microscopy

Topographical and electrochemical nanoscale imaging of living cells using voltage-switching mode scanning electrochemical microscopy

High Sensitive Elemental Analysis of Single Yeast Cells (Saccharomyces cerevisiae) by Time-Resolved Inductively-Coupled Plasma Mass Spectrometry Using a High Efficiency Cell Introduction System

Time-resolved ICP-MS Measurement: a New Method for Elemental and Multiparametric Analysis of Single Cells

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