Scanning Electrochemical Microscopy of Model Neurons:  Constant Distance Imaging

Kurulugama, Ruwan T.; Wipf, David O.; Takacs, Sara A.; Pongmayteegul, Sirinun; Garris, Paul A.; Baur, John E.

doi:10.1021/ac048571n

Cited by 147 publications

(130 citation statements)

References 44 publications

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“…In this way, the entire cell or a major portion of it can be imaged with a high spatial resolution by using a tip radius much smaller that the cell dimensions. By using this approach, high-quality topographic images of neuron cells were obtained with a spatial resolution of Ϸ1 m (14). Here, we used a small and very sharp tip with r g Ͻ 3a to increase the resolution to Ϸ200 nm.…”

Section: Discussionmentioning

confidence: 99%

“…The applications ranged from dopamine (4) and cholesterol (9) analysis to studies of menadione metabolism in yeast (10) and hepatoblastoma cells (11) to detection of reactive oxygen species (12) and monitoring the drug efflux from cancer cells (13). The SECM was also used for micrometer-scale topographic and reaction rate imaging of various types of biological cells (14)(15)(16)(17).…”

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confidence: 99%

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Nanoelectrochemistry of mammalian cells

Sun

Laforge

Abeyweera

et al. 2008

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

207

200

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There is a significant current interest in development of new techniques for direct characterization of the intracellular redox state and high-resolution imaging of living cells. We used nanometersized amperometric probes in combination with the scanning electrochemical microscope (SECM) to carry out spatially resolved electrochemical experiments in cultured human breast cells. With the tip radius Ϸ1,000 times smaller than that of a cell, an electrochemical probe can penetrate a cell and travel inside it without apparent damage to the membrane. The data demonstrate the possibility of measuring the rate of transmembrane charge transport and membrane potential and probing redox properties at the subcellular level. The same experimental setup was used for nanoscale electrochemical imaging of the cell surface.charge transfer ͉ membrane potential ͉ scanning electrochemical microscopy ͉ voltammetry

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Nanoelectrochemistry of mammalian cells

Sun

Laforge

Abeyweera

et al. 2008

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

207

200

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“…The reliable fabrication of nanoelectrodes with a small ratio of electrodeinsulation to active electrode (RG) is of particular importance to improve SECM spatial resolution (2). In particular, a variety of different approaches have been adopted to create small electrodes with thin insulating coats such as photolithography (3), chemical vapor deposition (4), electrodeposited paint methods (5,6), laser pulling techniques (7)(8)(9)(10)(11), and pyrolytic carbon deposition (12)(13)(14)(15). Photolithography and chemical vapor deposition can be used to make thin insulation layers, but pinholes are often a problem.…”

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confidence: 99%

“…On the other hand, pyrolytic carbon deposition inside glass pipettes enables fabrication of nano-sized electrodes with insulation of excellent integrity. This type of electrode has proven powerful for living cell measurements (14,15), and we thus use it for the studies herein.…”

mentioning

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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“…A constant-distance mode, in which the tip traces the contours of the surface based on the tip current, has been used [14], but there is the issue of how to separate unambiguously the topography and activity of the sample; a problem which can be addressed by implementing a voltage switching mode [15]. Another approach has been to integrate an UME into a tip for atomic force microscopy (AFM), in combined SECM-AFM [16][17][18][19][20], with high resolution tips having been made using nanowires grown on single-walled carbon nanotubes [21].…”

Section: Introductionmentioning

confidence: 99%

Nanoscale intermittent contact-scanning electrochemical microscopy

Lazenby

McKelvey

Peruffo

et al. 2013

J Solid State Electrochem

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Contribution to the Festschrift for Allen J. Bard, on the occasion of his 80 th birthday.2 Abstract A major theme in scanning electrochemical microscopy (SECM) is methodology for nanoscale imaging with distance control and positional feedback of the tip. We report the expansion of intermittent contact (IC)-SECM to the nanoscale, using disk-type Pt nanoelectrodes prepared using the laser-puller sealing method. The Pt was exposed using a focused ion beam milling procedure to cut the end of the electrode to a well-defined glass sheath radius, which could also be used to reshape the tips to reduce the size of the glass sheath. This produced nanoelectrodes that were slightly recessed, which was optimal for IC-SECM on the nanoscale, as it served to protect the active part of the tip. A combination of finite element method simulations, steady-state voltammety and scanning electron microscopy, for the measurement of critical dimensions, was used to estimate Pt recession depth. With this knowledge, the tip-substrate alignment could be further estimated by tip approach curve measurements. IC-SECM has been implemented by using a piezo-bender actuator for the detection of damping of the oscillation amplitude of the tip, when IC occurs, which was used as a tip position feedback mechanism. The piezo-bender actuator improves significantly on the performance of our previous setup for IC-SECM, as the force acting on the sample due to the tip is greatly reduced, allowing studies with more delicate tips. The capability of IC-SECM is illustrated with studies of a model electrode (metal/glass) substrate.

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Scanning Electrochemical Microscopy of Model Neurons: Constant Distance Imaging

Cited by 147 publications

References 44 publications

Nanoelectrochemistry of mammalian cells

Nanoelectrochemistry of mammalian cells

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

Nanoscale intermittent contact-scanning electrochemical microscopy

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