F. Picollo

Fabrication and electrical characterization of three-dimensional graphitic microchannels in single crystal diamond

We report on the systematic characterization of conductive micro-channels fabricated in single-crystal diamond with direct ion microbeam writing. Focused high-energy ( MeV) helium ions are employed to selectively convert diamond with micrometric spatial accuracy to a stable graphitic phase upon thermal annealing, due to the induced structural damage occurring at the end-of-range. A variable-thickness mask allows the accurate modulation of the depth at which the microchannels are formed, from several µm deep up to the very surface of the sample. By means of cross-sectional transmission electron microscopy (TEM), we demonstrate that the technique allows the direct writing of amorphous (and graphitic, upon suitable thermal annealing) microstructures extending within the insulating diamond matrix in the three spatial directions, and in particular, that buried channels embedded in a highly insulating matrix emerge and electrically connect to the sample surface at specific locations. Moreover, by means of electrical characterization at both

Direct fabrication of three-dimensional buried conductive channels in single crystal diamond with ion microbeam induced graphitization

Abstract We report on a novel method for the fabrication of three-dimensional buried graphitic micropaths in single crystal diamond with the employment of focused MeV ions. The use of implantation masks with graded thickness at the sub-micrometer scale allows the formation of conductive channels which are embedded in the insulating matrix at controllable depths. In particular, the modulation of the channels depth at their endpoints allows the surface contacting of the channel terminations with no need of further fabrication stages. In the present work we describe the sample masking, which includes the deposition of semi-spherical gold contacts on the sample surface, followed by MeV ion implantation. Because of the significant difference between the densities of pristine and amorphous or graphitized diamond, the formation of buried channels has a relevant mechanical effect on the diamond structure, causing localized surface swelling, which has been measured both with interferometric profilometry and atomic force microscopy. The electrical properties of the buried channels are then measured with a two point probe station: clear evidence is given that only the terminal points of the channels are electrically connected with the surface, while the rest of the channels extends below the surface. IV measurements are employed also to qualitatively investigate the electrical properties of the channels as a function of implantation fluence and annealing.

A New Diamond Biosensor with Integrated Graphitic Microchannels for Detecting Quantal Exocytic Events from Chromaffin Cells

An MeV ion-microbeam lithographic technique can be successfully employed for the fabrication of an all-carbon miniaturized cellular biosensor based on graphitic microchannels embedded in a single-crystal diamond matrix. The device is functionally characterized for the in vitro recording of quantal exocytic events from single chromaffin cells, with high sensitivity and signal-to-noise ratio, opening promising perspectives for the realization of monolithic all-carbon cellular biosensors.

Development and Characterization of a Diamond-Insulated Graphitic Multi Electrode Array Realized with Ion Beam Lithography

The detection of quantal exocytic events from neurons and neuroendocrine cells is a challenging task in neuroscience. One of the most promising platforms for the development of a new generation of biosensors is diamond, due to its biocompatibility, transparency and chemical inertness. Moreover, the electrical properties of diamond can be turned from a perfect insulator into a conductive material (resistivity ∼mΩ·cm) by exploiting the metastable nature of this allotropic form of carbon. A 16-channels MEA (Multi Electrode Array) suitable for cell culture growing has been fabricated by means of ion implantation. A focused 1.2 MeV He+ beam was scanned on a IIa single-crystal diamond sample (4.5 × 4.5 × 0.5 mm3) to cause highly damaged sub-superficial structures that were defined with micrometric spatial resolution. After implantation, the sample was annealed. This process provides the conversion of the sub-superficial highly damaged regions to a graphitic phase embedded in a highly insulating diamond matrix. Thanks to a three-dimensional masking technique, the endpoints of the sub-superficial channels emerge in contact with the sample surface, therefore being available as sensing electrodes. Cyclic voltammetry and amperometry measurements of solutions with increasing concentrations of adrenaline were performed to characterize the biosensor sensitivity. The reported results demonstrate that this new type of biosensor is suitable for in vitro detection of catecholamine release.

Formation of buried conductive micro-channels in single crystal diamond with MeV C and He implantation

Abstract As demonstrated in previous works, implantation with a MeV ion microbeam through masks with graded thickness allows the formation of conductive micro-channels in diamond which are embedded in the insulating matrix at controllable depths [P. Olivero et al., Diamond Relat. Mater. 18 (5–8), 870–876 (2009)]. In the present work we report about the systematic electrical characterization of such micro-channels as a function of several implantation conditions, namely: ion species and energy, implantation fluence. The current–voltage (IV) characteristics of the buried channels were measured at room temperature with a two point probe station. Significant parameters such as the sheet resistance and the characteristic exponent (α) of the IV power-law trend were expressed as a function of damage density, with satisfactory compatibility between the results obtained in different implantation conditions.

All-carbon multi-electrode array for real-time in vitro measurements of oxidizable neurotransmitters

We report on the ion beam fabrication of all-carbon multi electrode arrays (MEAs) based on 16 graphitic micro-channels embedded in single-crystal diamond (SCD) substrates. The fabricated SCD-MEAs are systematically employed for the in vitro simultaneous amperometric detection of the secretory activity from populations of chromaffin cells, demonstrating a new sensing approach with respect to standard techniques. The biochemical stability and biocompatibility of the SCD-based device combined with the parallel recording of multi-electrodes array allow: i) a significant time saving in data collection during drug screening and/or pharmacological tests over a large number of cells, ii) the possibility of comparing altered cell functionality among cell populations, and iii) the repeatition of acquisition runs over many cycles with a fully non-toxic and chemically robust bio-sensitive substrate.

Planar Diamond-Based Multiarrays to Monitor Neurotransmitter Release and Action Potential Firing: New Perspectives in Cellular Neuroscience

High biocompatibility, outstanding electrochemical responsiveness, inertness, and transparency make diamond-based multiarrays (DBMs) first-rate biosensors for in vitro detection of electrochemical and electrical signals from excitable cells together, with potential for in vivo applications as neural interfaces and prostheses. Here, we will review the electrochemical and physical properties of various DBMs and how these devices have been employed for recording released neurotransmitter molecules and all-or-none action potentials from living cells. Specifically, we will overview how DBMs can resolve localized exocytotic events from subcellular compartments using high-density microelectrode arrays (MEAs), or monitoring oxidizable neurotransmitter release from populations of cells in culture and tissue slices using low-density MEAs. Interfacing DBMs with excitable cells is currently leading to the promising opportunity of recording electrical signals as well as creating neuronal interfaces through the same device. Given the recent increasingly growing development of newly available DBMs of various geometries to monitor electrical activity and neurotransmitter release in a variety of excitable and neuronal tissues, the discussion will be limited to planar DBMs.

Effects of high-power laser irradiation on sub-superficial graphitic layers in single-crystal diamond

We report on the structural modifications induced by a λ = 532 nm ns-pulsed high-power laser on sub-superficial graphitic layers in single-crystal diamond realized by means of MeV ion implantation. A systematic characterization of the structures obtained under different laser irradiation conditions (power density, number of pulses) and subsequent thermal annealing was performed by different electron microscopy techniques. The main feature observed after laser irradiation is the thickening of the pre-existing graphitic layer. Cross-sectional SEM imaging was performed to directly measure the thickness of the modified layers, and subsequent selective etching of the buried layers was employed to both assess their graphitic nature and enhance the SEM imaging contrast. In particular, it was found that for optimal irradiation parameters the laser processing induces a six-fold increase the thickness of sub-superficial graphitic layers without inducing mechanical failures in the surrounding crystal. TEM microscopy and EELS spectroscopy allowed a detailed analysis of the internal structure of the laser-irradiated layers, highlighting the presence of different nano-graphitic and amorphous layers. The obtained results demonstrate the effectiveness and versatility of high-power laser irradiation for an accurate tuning of the geometrical and structural features of graphitic structures embedded in single-crystal diamond, and open new opportunities in diamond fabrication.

Nanodiamonds-induced effects on neuronal firing of mouse hippocampal microcircuits

Fluorescent nanodiamonds (FND) are carbon-based nanomaterials that can efficiently incorporate optically active photoluminescent centers such as the nitrogen-vacancy complex, thus making them promising candidates as optical biolabels and drug-delivery agents. FNDs exhibit bright fluorescence without photobleaching combined with high uptake rate and low cytotoxicity. Focusing on FNDs interference with neuronal function, here we examined their effect on cultured hippocampal neurons, monitoring the whole network development as well as the electrophysiological properties of single neurons. We observed that FNDs drastically decreased the frequency of inhibitory (from 1.81 Hz to 0.86 Hz) and excitatory (from 1.61 to 0.68 Hz) miniature postsynaptic currents, and consistently reduced action potential (AP) firing frequency (by 36%), as measured by microelectrode arrays. On the contrary, bursts synchronization was preserved, as well as the amplitude of spontaneous inhibitory and excitatory events. Current-clamp recordings revealed that the ratio of neurons responding with AP trains of high-frequency (fast-spiking) versus neurons responding with trains of low-frequency (slow-spiking) was unaltered, suggesting that FNDs exerted a comparable action on neuronal subpopulations. At the single cell level, rapid onset of the somatic AP (“kink”) was drastically reduced in FND-treated neurons, suggesting a reduced contribution of axonal and dendritic components while preserving neuronal excitability.

Direct experimental observation of nonclassicality in ensembles of single-photon emitters

In this work we experimentally demonstrate a recently proposed criterion addressed to detect nonclassical behavior in the fluorescence emission of ensembles of single-photon emitters. In particular, we apply the method to study clusters of nitrogen-vacancy centers in diamond characterized with single-photon-sensitive confocal microscopy. Theoretical considerations on the behavior of the parameter at any arbitrary order in the presence of Poissonian noise are presented and, finally, the opportunity of detecting manifold coincidences is discussed.