Towards a planar sample support for in situ experiments in structural biology

Gullo, Maurizio R.; Akiyama, T.; Frederix, P. L. T. M.; Tonin, A.; Staufer, U.; Engel, Andréas; Rooij, N. F. de

doi:10.1016/j.mee.2004.12.072

Cited by 5 publications

(1 citation statement)

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“…The set-up for conducting experiments on cell-membrane patches was to be completed with a planar patch-clamp like sample support [6]. Such a device will allow electronically accessing both sides of the sample: On one side, a global electrode will be used, where as on the other side, the conductive AFM tip will be employed to locally apply a potential.…”

Section: Nanotools For Biologymentioning

confidence: 99%

Micro- and nanosystems for biology and medicine

Staufer

Akiyama

Gullo

et al. 2007

Microelectronic Engineering

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The development of new tools and instruments for biomedical applications based on nano-(NEMS) or microelectromechanical systems technology (MEMS) are bridging the gap between the macro-and the nano-world. The well mastered microtechnique allows controlling many parameters of these instruments, which is essential for conducting reproducible and repeatable experiments in the life sciences. Examples are multifunctional scanning probe sensors for cell biology, an arthroscopic scanning force microscope for minimally invasive medical interventions and a nanopore sensor for single molecule experiments in biochemistry. This paper reviews some of the activities conducted in a fruitful interdisciplinary collaboration between physicists, engineers, biologists and physicians.Keywords: Microsystems; Nanosystems; Cell biology; AFM; Nanopore; Protein; Cartilage; Minimal invasive instrument Nanotools for biologyIn cell and structural biology the typical dimensions are spanning the length scales from a few nanometers up to centimeters. The objects of interest are very delicate and often mechanically soft, hence, investigating their functions calls for gentle and precise instruments. Very often several parameters should be simultaneously controlled in such experiments and a more or less complex system of tools is required. An appealing implementation of such systems is to employ micro-and nanofabrication techniques.Plain atomic force microscope [AFM] probes have proven to provide images of membrane proteins with subnanometer resolution in the past [1]. In order to study structural changes in voltage gated channel proteins at this level of resolution, we have developed a probe, which features a conductive tip [2]. The metal is insulated up to the apex (Fig. 1) such that Faradaic currents in buffer solutions are minimal. Different metals were tested, and PtSi proofed to be the best choice from a fabrication point of view [3]. The basic concept for fabricating such a probe followed that of standard molded silicon nitride tips. We employed anisotropic KOH etching of silicon (1 0 0)-wafers through a silicon nitride mask in order to form small pyramidal etch-pits. In one of these pits the tip will be formed. An array of them will be used as contact area. All pits were sharpened by a local low-temperature oxidation process, followed by a deposition of a thin layer of polycrystalline Si. The latter was lithographically patterned such that a thin line connection between the tip mold and the array of pits was formed. Platinum was evaporated on the full wafer and then annealed in order to form the silicide. Residual Pt was removed, and a second silicon nitride layer was deposited by means of low pressure chemical vapor deposition [LPCVD] in order to fully encapsulate the metal structure. The cantilevers were patterned by lithography and reactive ion etching [RIE], a pre-structured Pyrex-glass wafer was bonded to the nitride, and the silicon substrate was dissolved in KOH. The final step was a timed etching in buffered hydrofluoric ...

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Section: Nanotools For Biologymentioning

confidence: 99%

Micro- and nanosystems for biology and medicine

Staufer

Akiyama

Gullo

et al. 2007

Microelectronic Engineering

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Planar Patch Clamp for Neuronal Networks—Considerations and Future Perspectives

Bosca

Martina

2014

Methods in Molecular Biology

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The patch-clamp technique is generally accepted as the gold standard for studying ion channel activity allowing investigators to either "clamp" membrane voltage and directly measure transmembrane currents through ion channels, or to passively monitor spontaneously occurring intracellular voltage oscillations. However, this resulting high information content comes at a price. The technique is labor-intensive and requires highly trained personnel and expensive equipment. This seriously limits its application as an interrogation tool for drug development. Patch-clamp chips have been developed in the last decade to overcome the tedious manipulations associated with the use of glass pipettes in conventional patch-clamp experiments. In this chapter, we describe some of the main materials and fabrication protocols that have been developed to date for the production of patch-clamp chips. We also present the concept of a patch-clamp chip array providing high resolution patch-clamp recordings from individual cells at multiple sites in a network of communicating neurons. On this chip, the neurons are aligned with the aperture-probes using chemical patterning. In the discussion we review the potential use of this technology for pharmaceutical assays, neuronal physiology and synaptic plasticity studies.

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Planar Patch Clamping

Behrends

Fertig

2007

Neuromethods

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Towards a planar sample support for in situ experiments in structural biology

Cited by 5 publications

References 10 publications

Micro- and nanosystems for biology and medicine

Micro- and nanosystems for biology and medicine

Planar Patch Clamp for Neuronal Networks—Considerations and Future Perspectives

Planar Patch Clamping

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