Microcantilever-based sensing arrays for evaluation of biomolecular interactions

The controlled immobilization on a surface of biomolecules used as recognition elements is of fundamental importance in order to realize highly specific and sensible biosensors. Microcantilevers (MC) are nanomechanical sensors, which can be used as label free micro-sized mechanical transducers. MC resonant frequency is sensitively modified upon molecules adsorption, demonstrating an impressive mass resolution. A widely used approach for the immobilization of biorecognition elements on silicon substrates consists in the deposition of 3-aminopropyl-triethoxysilane (APTES) followed by the incubation with glutaraldehyde (GA) as a crosslinking agent. However, these derivatization processes produce a variable chemical functionalization because of the spontaneous polymerization of GA in aqueous solutions. With the aim of producing a more reliable chemical functionalization for protein immobilization, the deposition of a thin film of APTES by self-assembly followed by the modification of its amino groups into carboxyl groups by incubating in succinic anhydride (SA) is proposed. Moreover, the activation of these terminal carboxyl groups were performed by using the EDC/s-NHS protocol in order to enhance their reactivity toward primary amine groups present on biomolecules surface. This method was characterized from a physico-chemical point of view by means of compositional and morphological surface analysis. Moreover, data acquired after the application of this functionalization to a MC-based system showed a highly reproducible deposition of APTES/SA when compared to APTES/GA deposition process. APTES/SA derivatized MC arrays were then incubated with biomolecules for the study of its protein binding capability: the quantification of the grafted biomolecules was performed from the gravimetric data and compared with a theoretical surface density calculated through a molecular modeling tool, providing information about the orientation of the proteins tethered to the surface. In order to avoid or reduce non-specific protein interactions, Bovine Serum Albumin and ethanolamine were considered for their blocking capability. Finally, the detection of the envelope glycoprotein domain III of the Dengue virus type 1 based on immune-specific recognition through the DV32.6 antibody was performed, providing a stoichiometry ratio for the DIII-DV1/DV32.6 interaction. Currently, no cure or vaccine are available; thus, a better understanding of the interactions between the viruses and specific antibodies is expected to provide fundamental information for the development of a vaccine

Morphological Characterization and Lumped Element Model of Graphene and Biochar Thick Films

Carbon based materials exhibit interesting mechanical, thermal and electrical properties which make them excellent contenders for use as fillers in composites as film. Graphene has been vastly used among the carbon-based materials. More recently eco-friendly carbon-based materials like biochar have emerged. The deployment of carbon-based materials in films needs to be studied since films are more versatile and permit the exploitation of electrical properties of such materials over circuits and systems. Typical circuits and systems exploiting electrical properties of novel materials perform a number of applications including sensing, detection, tunable devices and energy harvesting. In this paper, films composed of 9:1 graphene or biochar are deployed on a microstrip line. The morphological properties of graphene and biochar and their respective films are studied with Raman spectra and Field Emission Scanning Electron Microscope (FESEM). The electrical properties (four-point probe measurements and scattering parameter measurements) of the films. Low frequency measurements are used as starting point for circuit models estimating the lumped impedance of the films. From the morphological characterization it is shown that biochar films appear as granulates carbonaceous materials whereas graphene films contains several flakes forming a network. From the low frequency measurements and microwave characterization it is seen that graphene films are more conductive as compared to biochar films. In many applications, it is useful to know the surface impedance of the film since it varies on interaction with any external stimulus (variation of pressure, humidity, gas, etc.)

Design of a Portable Microfluidic Platform for EGOT-Based in Liquid Biosensing

In biosensing applications, the exploitation of organic transistors gated via a liquid electrolyte has increased in the last years thanks to their enormous advantages in terms of sensitivity, low cost and power consumption. However, a practical aspect limiting the use of these devices in real applications is the contamination of the organic material, which represents an obstacle for the realization of a portable sensing platform based on electrolyte-gated organic transistors (EGOTs). In this work, a novel contamination-free microfluidic platform allowing differential measurements is presented and validated through finite element modeling simulations. The proposed design allows the exposure of the sensing electrode without contaminating the EGOT device during the whole sensing tests protocol. Furthermore, the platform is exploited to perform the detection of bovine serum albumin (BSA) as a validation test for the introduced differential protocol, demonstrating the capability to detect BSA at 1 pM concentration. The lack of contamination and the differential measurements provided in this work can be the first steps towards the realization of a reliable EGOT-based portable sensing instrument

Biomedical Applications

In recent years, 3D printing technology has become a sufficiently mature technique to allow not only the production of objects starting from a design modeling but also a possible customization through the introduction of functionality by the end user. This rapid prototyping technique represents a very promising technology for device fabrication with different application fields (e.g., biological, environmental, food, aerospace), offering advantages over traditional manufacturing methods. Moreover, the 3D printing archetype has introduced novel opportunities for the realization of smart devices, where the added value lies in their intrinsic functionality. In fact, 3D-printed functional materials can transform their structure in response to specific stimuli (e.g., temperature, pH, light radiation, etc.), adding to the printed object interesting properties to take advantage of. Recently, this paradigm has been explored and expanded by researcher’s/engineer’s community with the aim of realizing 3D printable objects that present exploitable chemical functionalities. In this chapter, functional 3D objects for bio-applications have been reported, combining 3D printing technology with an accurate material engineering. The result is a single step 3D-printed object with intrinsic chemical functionalities that could be exploited to produce immunoassay-based and/or enzymatically active devices for biosensing purposes and precision medicine

Experimental evidence of Fano resonances in nanomechanical resonators

Fano resonance refers to an interference between localized and continuum states that was firstly reported for atomic physics and solid-state quantum devices. In recent years, Fano interference gained more and more attention for its importance in metamaterials, nanoscale photonic devices, plasmonic nanoclusters and surface-enhanced Raman scattering (SERS). Despite such interest in nano-optics, no experimental evidence of Fano interference was reported up to now for purely nanomechanical resonators, even if classical mechanical analogies were referred from a theoretical point of view. Here we demonstrate for the first time that harmonic nanomechanical resonators with relatively high quality factors, such as cantilevers vibrating in vacuum, can show characteristic Fano asymmetric curves when coupled in arrays. The reported findings open new perspectives in fundamental aspects of classical nanomechanical resonators and pave the way to a new generation of chemical and biological nanoresonator sensors with higher parallelization capability

Optimization and characterization of a homogeneous carboxylic surface functionalization for silicon-based biosensing

A well-organized immobilization of bio-receptors is a crucial goal in biosensing, especially to achieve high reproducibility, sensitivity and specificity. These requirements are usually attained with a controlled chemical/biochemical functionalization that creates a stable layer on a sensor surface. In this work, a chemical modification protocol for silicon-based surfaces to be applied in biosensing devices is presented. An anhydrous silanization step through 3-aminopropylsilane (APTES), followed by a further derivatization with succinic anhydride (SA), is optimized to generate an ordered flat layer of carboxylic groups. The properties of APTES/SA modified surface were compared with a functionalization in which glutaraldehyde (GA) is used as crosslinker instead of SA, in order to have a comparison with an established and largely applied procedure. Moreover, a functionalization based on the controlled deposition of a plasma polymerized acrylic acid (PPAA) thin film was used as a reference for carboxylic reactivity. Advantages and drawbacks of the considered methods are highlighted, through physico-chemical characterizations (OCA, XPS, and AFM) and by means of a functional Protein G/Antibody immunoassay. These analyses reveal that the most homogeneous, reproducible and active surface is achieved by using the optimized APTES/SA coupling