Diamond-based sensors for in vitro cellular radiobiology: Simultaneous detection of cell exocytic activity and ionizing radiation.

The investigation of secondary effects induced by ionizing radiation represents a new and ever-growing research field in radiobiology. This new paradigm cannot be investigated only using standard instrumentation and methodologies, but rather requires novel technologies to achieve significant progress. In this framework, we developed diamond-based sensors that allow simultaneous real-time measurements with a high spatial resolution of the secretory activity of a network of cells cultured on the device, as well as of the dose at which they are exposed during irradiation experiments. The devices were functionally characterized by testing both the above-mentioned detection schemes, namely: amperometric measurements of neurotransmitter release from excitable cells (such as dopamine or adrenaline) and dosimetric evaluation using different ionizing particles (alpha particle and X-ray photons). Finally, the sensors were employed to investigate the effects induced by X-rays on the exocytotic activity of PC12 neuroendocrine cells by monitoring the modulation of the dopamine release in real-time.

Spectroscopic measurement of the refractive index of ion-implanted diamond.

We present the results of variable-angle spectroscopic ellipsometry and transmittance measurements to determine the variation of the complex refractive index of ion-implanted single-crystal diamond. An increase is found in both real and imaginary parts at increasing damage densities. The index depth variation is determined in the whole wavelength range between 250 and 1690 nm. The dependence from the vacancy density is evaluated, highlighting a deviation from linearity in the high-damage-density regime. A considerable increase (up to 5%) in the real part of the index is observed, attributed to an increase in polarizability, thus offering new microfabrication possibilities for waveguides and other photonic structures in diamond.

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.

Laser-induced etching of few-layer graphene synthesized by Rapid-Chemical Vapour Deposition on Cu thin films.

The outstanding electrical and mechanical properties of graphene make it very attractive for several applications, Nanoelectronics above all. However a reproducible and non destructive way to produce high quality, large-scale area, single layer graphene sheets is still lacking. Chemical Vapour Deposition of graphene on Cu catalytic thin films represents a promising method to reach this goal, because of the low temperatures (T < 950°C-1000°C) involved during the process and of the theoretically expected monolayer self-limiting growth. On the contrary such self-limiting growth is not commonly observed in experiments, thus making the development of techniques allowing for a better control of graphene growth highly desirable. Here we report about the local ablation effect, arising in Raman analysis, due to the heat transfer induced by the laser incident beam onto the graphene sample.

A diamond-based biosensor for the recording of neuronal activity.

We have developed a device for recording the extracellular electrical activity of cultured neuronal networks based on a hydrogen terminated (H-terminated) conductive diamond. GT1-7 cells, a neuronal cell line showing spontaneous action potentials firing, could maintain their functional properties for days in culture when plated on the H-terminated diamond surface. The recorded extracellular electrical activity appeared in the form of well-resolved bursts of fast and slow biphasic signals with a mean duration of about 8ms for the fast and 60ms for the slow events. The time courses of these signals were in good agreement with those recorded by means of conventional microelectrode array (MEAs) and with the negative derivative of the action potentials intracellularly recorded with the patch clamp technique from single cells. Thus, although hydrophobic in nature, the conductive H-terminated diamond surface is able to reveal the spontaneous electrical activity of neurons mainly by capacitative coupling to the cell membrane. Having previously shown that the optical properties of H-terminated diamond allow to record cellular activity by means of fluorescent probes (Ariano, P., Baldelli, P., Carbone, E., Giardino, A., Lo Giudice, A., Lovisolo, D., Manfredotti, C., Novara, M., Sternschulte, H., Vittone, E., 2005. Diam. Relat. Mater. 14, 669-674), we now provide evidence for the feasibility of using diamond-based cellular biosensors for multiparametrical recordings of electrical activity from living cells.