48-spot single-molecule FRET setup with periodic acceptor excitation.

Single-molecule Förster resonance energy transfer (smFRET) allows measuring distances between donor and acceptor fluorophores on the 3-10 nm range. Solution-based smFRET allows measurement of binding-unbinding events or conformational changes of dye-labeled biomolecules without ensemble averaging and free from surface perturbations. When employing dual (or multi) laser excitation, smFRET allows resolving the number of fluorescent labels on each molecule, greatly enhancing the ability to study heterogeneous samples. A major drawback to solution-based smFRET is the low throughput, which renders repetitive measurements expensive and hinders the ability to study kinetic phenomena in real-time. Here we demonstrate a high-throughput smFRET system that multiplexes acquisition by using 48 excitation spots and two 48-pixel single-photon avalanche diode array detectors. The system employs two excitation lasers allowing separation of species with one or two active fluorophores. The performance of the system is demonstrated on a set of doubly labeled double-stranded DNA oligonucleotides with different distances between donor and acceptor dyes along the DNA duplex. We show that the acquisition time for accurate subpopulation identification is reduced from several minutes to seconds, opening the way to high-throughput screening applications and real-time kinetics studies of enzymatic reactions such as DNA transcription by bacterial RNA polymerase.

Silicon photon-counting avalanche diodes for single-molecule fluorescence spectroscopy.

Solution-based single-molecule fluorescence spectroscopy is a powerful experimental tool with applications in cell biology, biochemistry and biophysics. The basic feature of this technique is to excite and collect light from a very small volume and work in a low concentration regime resulting in rare burst-like events corresponding to the transit of a single molecule. Detecting photon bursts is a challenging task: the small number of emitted photons in each burst calls for high detector sensitivity. Bursts are very brief, requiring detectors with fast response time and capable of sustaining high count rates. Finally, many bursts need to be accumulated to achieve proper statistical accuracy, resulting in long measurement time unless parallelization strategies are implemented to speed up data acquisition. In this paper we will show that silicon single-photon avalanche diodes (SPADs) best meet the needs of single-molecule detection. We will review the key SPAD parameters and highlight the issues to be addressed in their design, fabrication and operation. After surveying the state-of-the-art SPAD technologies, we will describe our recent progress towards increasing the throughput of single-molecule fluorescence spectroscopy in solution using parallel arrays of SPADs. The potential of this approach is illustrated with single-molecule Förster resonance energy transfer measurements.

Impact of Surface States in Graphene/p-Si Schottky Diodes.

Graphene-silicon Schottky diodes are intriguing devices that straddle the border between classical models and two-dimensional ones. Many papers have been published in recent years studying their operation based on the classical model developed for metal-silicon Schottky diodes. However, the results obtained for diode parameters vary widely in some cases showing very large deviations with respect to the expected range. This indicates that our understanding of their operation remains incomplete. When modeling these devices, certain aspects strictly connected with the quantum mechanical features of both graphene and the interface with silicon play a crucial role and must be considered. In particular, the dependence of the graphene Fermi level on carrier density, the relation of the latter with the density of surface states in silicon and the coupling between in-plane and out-of-plane dynamics in graphene are key aspects for the interpretation of their behavior. Within the thermionic regime, we estimate the zero-bias Schottky barrier height and the density of silicon surface states in graphene/type-p silicon diodes by adapting a kown model and extracting ideality index values close to unity. The ohmic regime, beyond the flat band potential, is modeled with an empirical law, and the current density appears to be roughly proportional to the electric field at the silicon interface; moreover, the graphene-to-silicon electron tunneling efficiency drops significantly in the transition from the thermionic to ohmic regime. We attribute these facts to (donor) silicon surface states, which tend to be empty in the ohmic regime.

The Physics behind the Modulation of Thermionic Current in Photodetectors Based on Graphene Embedded between Amorphous and Crystalline Silicon.

In this work, we investigate a vertically illuminated near-infrared photodetector based on a graphene layer physically embedded between a crystalline and a hydrogenated silicon layer. Under near-infrared illumination, our devices show an unforeseen increase in the thermionic current. This effect has been ascribed to the lowering of the graphene/crystalline silicon Schottky barrier as the result of an upward shift in the graphene Fermi level induced by the charge carriers released from traps localized at the graphene/amorphous silicon interface under illumination. A complex model reproducing the experimental observations has been presented and discussed. Responsivity of our devices exhibits a maximum value of 27 mA/W at 1543 nm under an optical power of 8.7 µW, which could be further improved at lower optical power. Our findings offer new insights, highlighting at the same time a new detection mechanism which could be exploited for developing near-infrared silicon photodetectors suitable for power monitoring applications.

Electrical conduction and noise spectroscopy of sodium-alginate gold-covered ultrathin films for flexible green electronics.

Green electronics is an emerging topic that requires the exploration of new methodologies for the integration of green components into electronic devices. Therefore, the development of alternative and eco-friendly raw materials, biocompatible and biodegradable, is of great importance. Among these, sodium-alginate is a natural biopolymer extracted from marine algae having a great potential in terms of transparency, flexibility, and conductivity, when functionalized with a thin gold (Au) layer. The electrical transport of these flexible and conducting substrates has been studied, by DC measurements, from 300 to 10 K, to understand the interplay between the organic substrate and the metallic layer. The results were compared to reference bilayers based on polymethyl-methacrylate, a well-known polymer used in electronics. In addition, a detailed investigation of the electric noise properties was also performed. This analysis allows to study the effect of charge carriers fluctuations, providing important information to quantify the minimum metallic thickness required for electronic applications. In particular, the typical noise behavior of metallic compounds was observed in samples covered with 5 nm of Au, while noise levels related to a non-metallic conduction were found for a thickness of 4.5 nm, despite of the relatively good DC conductance of the bilayer.

Electric Transport in Gold-Covered Sodium-Alginate Free-Standing Foils.

The electric transport properties of flexible and transparent conducting bilayers, realized by sputtering ultrathin gold nanometric layers on sodium-alginate free-standing films, were studied. The reported results cover a range of temperatures from 3 to 300 K. In the case of gold layer thicknesses larger than 5 nm, a typical metallic behavior was observed. Conversely, for a gold thickness of 4.5 nm, an unusual resistance temperature dependence was found. The dominant transport mechanism below 70 K was identified as a fluctuation-induced tunneling process. This indicates that the conductive region is not continuous but is formed by gold clusters embedded in the polymeric matrix. Above 70 K, instead, the data can be interpreted using a phenomenological model, which assumes an anomalous expansion of the conductive region upon decreasing the temperature, in the range from 300 to 200 K. The approach herein adopted, complemented with other characterizations, can provide useful information for the development of innovative and green optoelectronics.

Optical crosstalk in SPAD arrays for high-throughput single-molecule fluorescence spectroscopy.

Single-molecule fluorescence spectroscopy (SMFS), based on the detection of individual molecules freely diffusing through the excitation spot of a confocal microscope, has allowed unprecedented insights into biological processes at the molecular level, but suffers from limited throughput. We have recently introduced a multispot version of SMFS, which allows achieving high-throughput SMFS by virtue of parallelization, and relies on custom silicon single-photon avalanche diode (SPAD) detector arrays. Here, we examine the premise of this parallelization approach, which is that data acquired from different spots is uncorrelated. In particular, we measure the optical crosstalk characteristics of the two 48-pixel SPAD arrays used in our recent SMFS studies, and demonstrate that it is negligible (crosstalk probability ≤ 1.1 10-3) and undetectable in cross-correlation analysis of actual single-molecule fluorescence data.

New silicon technologies enable high-performance arrays of Single Photon Avalanche Diodes.

In order to fulfill the requirements of many applications, we recently developed a new technology aimed at combining the advantages of traditional thin and thick silicon Single Photon Avalanche Diodes (SPAD). In particular we demonstrated single-pixel detectors with a remarkable improvement in the Photon Detection Efficiency at the longer wavelengths (e.g. 40% at 800nm) while maintaining a timing jitter better than 100ps. In this paper we will analyze the factors the currently prevent the fabrication of arrays of SPADs by adopting such a Red-Enhanced (RE) technology and we will propose further modifications to the device structure that will enable the fabrication of high performance RE-SPAD arrays for photon timing applications.

A 48-pixel array of Single Photon Avalanche Diodes for multispot Single Molecule analysis.

In this paper we present an array of 48 Single Photon Avalanche Diodes (SPADs) specifically designed for multispot Single Molecule Analysis. The detectors have been arranged in a 12×4 square geometry with a pitch-to-diameter ratio of ten in order to minimize the collection of the light from non-conjugated excitation spots. In order to explore the trade-offs between the detectors' performance and the optical coupling with the experimental setup, SPADs with an active diameter of 25 and of 50µm have been manufactured. The use of a custom technology, specifically designed for the fabrication of the detectors, allowed us to combine a high photon detection efficiency (peak close to 50% at a wavelength of 550nm) with a low dark count rate compatible with true single molecule detection. In order to allow easy integration into the optical setup for parallel single-molecule analysis, the SPAD array has been incorporated in a compact module containing all the electronics needed for a proper operation of the detectors.

Liquid-crystalline rigid-core semiconductor oligothiophenes: influence of molecular structure on phase behaviour and thin-film properties.

The design, synthesis and properties of liquid-crystalline semiconducting oligothiophenes containing dithienothiophene (DTT), benzothiadiazole (BTZ) and carbazole (CBZ) rigid cores are described. The effect of molecular structure (shape, size and substitution) on their thermal behaviour and electrical properties has been investigated. Polarised optical microscopy (POM) and differential scanning calorimetry (DSC) analyses have revealed highly ordered smectic mesophases for most of the newly synthesised compounds. X-ray diffraction (XRD) studies performed at various temperatures have shown that the smectic order is retained in the crystalline state upon cooling across the transition temperature, affording cast films with a more favourable morphology for FET applications.

Multiscale self-organization of the organic semiconductor alpha-quinquethiophene.

We show that thin films grown by vacuum sublimation, or formed by melted powders, of semiconductor alpha-quinquethiophene (T5) exhibit a hierarchical self-affinity organization that spans scales from tens of nanometers to hundreds of micrometers. T5 organization was investigated by X-ray diffraction (XRD), atomic force microscopy (AFM), and optical microscopy. XRD showed that vacuum-evaporated T5 films were characterized by a preferred orientation of the h00 planes parallel to the glass substrate. Melting of the films followed by rapid quenching to room temperature led to the formation of micrometer-sized, single-crystal-like structures, characterized by uniaxially aligned stripes. XRD proved that the melting-quenching process enhanced molecular ordering and increased the size of domains with the molecule's long axes tilted by about 65 degrees with respect to the substrate plane and piled up side-by-side along parallel columns. AFM measurements on the melt-quenched structures showed that a hierarchical architecture was built by reiteration across multiple length scales of the same recurring motif. Because of the tendency of T5 to form highly crystalline vacuum-evaporated thin films, a field-effect hole mobility comparable to state-of-the-art FET mobility of alpha-sexithiophene films was reached, without any attempt to optimize deposition conditions.