M. L. Coluccio

Microfluidics & nanotechnology: towards fully integrated analytical devices for the detection of cancer biomarkers

In this paper, we describe an innovative modular microfluidic platform allowing filtering, concentration and analysis of peptides from a complex mixture. The platform is composed of a microfluidic filtering device and a superhydrophobic surface integrating surface enhanced Raman scattering (SERS) sensors. The microfluidic device was used to filter specific peptides (MW 1553.73 D) derived from the BRCA1 protein, a tumor-suppressor molecule which plays a pivotal role in the development of breast cancers, from albumin (66.5 KD), the most represented protein in human plasma. The filtering process consisted of driving the complex mixture through a porous membrane having a cut-off of 12–14 kD by hydrodynamic flow. The filtered samples coming out of the microfluidic device were subsequently deposited on a superhydrophobic surface formed by micro pillars on top of which nanograins were fabricated. The nanograins coupled to a Raman spectroscopy instrument acted as a SERS sensor and allowed analysis of the filtered sample on top of the surface once it evaporated. By using the presented platform, we demonstrate being able to sort small peptides from bigger proteins and to detect them by using a label-free technique at a resolution down to 0.1 ng μL−1. The combination of microfluidics and nanotechnology to develop the presented microfluidic platform may give rise to a new generation of biosensors capable of detecting low concentration samples from complex mixtures without the need for any sample pretreatment or labelling. The developed devices could have future applications in the field of early diagnosis of severe illnesses, e.g. early cancer detection.

From nucleotides to DNA analysis by a SERS substrate of a self similar chain of silver nanospheres

In this work we realized a device of silver nanostructures designed so that they have a great ability to sustain the surface-enhanced Raman scattering effect. The nanostructures were silver self-similar chains of three nanospheres, having constant ratios between their diameters and between their reciprocal distances. They were realized by electron beam lithography, to write the pattern, and by silver electroless deposition technique, to fill it with the metal. The obtained device showed the capability to increase the Raman signal coming from the gap between the two smallest nanospheres (whose size is around 10 nm) and so it allows the detection of biomolecules fallen into this hot spot. In particular, oligonucleotides with 6 DNA bases, deposited on these devices with a drop coating method, gave a Raman spectrum characterized by a clear fingerprint coming from the hot spot and, with the help of a fitting method, also oligonucleotides of 9 bases, which are less than 3 nm long, were resolved. In conclusion the silver nanolens results in a SERS device able to measure all the molecules, or part of them, held into the hot spot of the nanolenses, and thus it could be a future instrument with which to analyze DNA portions.

Electroless formation of silver nanoaggregates: an experimental and molecular dynamics approach

The ability to manipulate matter to create non-conventional structures is one of the key issues of material science. The understanding of assembling mechanism at the nanoscale allows us to engineer new nanomaterials, with physical properties intimately depending on their structure. This paper describes new strategies to obtain and characterise metal nanostructures via the combination of a top-down method, such as electron beam lithography, and a bottom-up technique, such as the chemical electroless deposition. We realised silver nanoparticle aggregates within well-defined patterned holes created by electron beam lithography on silicon substrates. The quality characteristics of the nanoaggregates were verified by using scanning electron microscopy and atomic force microscopy imaging. Moreover, we compared the experimental findings to molecular dynamics simulations of nanoparticles growth. We observed a very high dependence of the structure characteristics on the pattern nanowell aspect ratio. We found that high-quality metal nanostructures may be obtained in patterns with well aspect ratio close to one, corresponding to a maximum diameter of 50 nm, a limit above which the fabricated structures become less regular and discontinuous. When regular shapes and sizes are necessary, as in nanophotonics, these results suggest the pattern characteristics to obtain isolated, uniform and reproducible metal nanospheres.

Silica diatom shells tailored with Au nanoparticles enable sensitive analysis of molecules for biological, safety and environment applications

Diatom shells are a natural, theoretically unlimited material composed of silicon dioxide, with regular patterns of pores penetrating through their surface. For their characteristics, diatom shells show promise to be used as low cost, highly efficient drug carriers, sensor devices or other micro-devices. Here, we demonstrate diatom shells functionalized with gold nanoparticles for the harvesting and detection of biological analytes (bovine serum albumin—BSA) and chemical pollutants (mineral oil) in low abundance ranges, for applications in bioengineering, medicine, safety, and pollution monitoring.

Nano-topography Enhances Communication in Neural Cells Networks

Neural cells are the smallest building blocks of the central and peripheral nervous systems. Information in neural networks and cell-substrate interactions have been heretofore studied separately. Understanding whether surface nano-topography can direct nerve cells assembly into computational efficient networks may provide new tools and criteria for tissue engineering and regenerative medicine. In this work, we used information theory approaches and functional multi calcium imaging (fMCI) techniques to examine how information flows in neural networks cultured on surfaces with controlled topography. We found that substrate roughness Sa affects networks topology. In the low nano-meter range, Sa = 0–30 nm, information increases with Sa. Moreover, we found that energy density of a network of cells correlates to the topology of that network. This reinforces the view that information, energy and surface nano-topography are tightly inter-connected and should not be neglected when studying cell-cell interaction in neural tissue repair and regeneration.

Plasmonics and Super-Hydrophobicity: A New Class of Nano-Bio-Devices

Early detection of diseases has great importance in terms of success of the disease treatment. In fact, it has a profound positive influence on the response provided by the patient, leading to shorter and less invasive treatment regimes. We consider here the Raman detection of low (atto-molar) concentrates of molecules by applying nanofabrication techniques in the fabrication of plasmonic devices fulfilling the requirement of superhydrophobicity. Plasmonic resonances will have the effect of substantially increasing the local electric field around the fabricated nano-device which, in turn, will positively affect the Raman signal. Similarly, the superhydrophobicity will play the crucial role in localizing the few molecules of the analyte around the plasmonic device, therefore allowing their detection in a manner otherwise impossible in diffusion-based devices. We will theoretically explain the concept of superhydrophobicity by providing also a roadmap for defining the optimal superhydrophobic device, then we will introduce the fabrication process to realize such a device and, finally, we will provide the Raman counting of a series of analytes together with electromagnetic simulations illustrating the role of the electric field in the formation of the Raman signal.

Adiabatic focusing of surface plasmon polaritons for label free detection of few molecules by means of Raman scattering

Here we report the design, the fabrication and measurement of a photonic-plasmonic device that is fully compatible with AFM microscopy and surface enhanced Raman spectroscopy. The physical mechanism exploited is the adiabatic compression of Surface Plasmon Polaritons which propagate along a silver nanocone generating a very high electric field at the tip end. A photonic crystal cavity is employed to efficiently couple the external laser radiation with the nanocone. The reported measurements demonstrate the accumulation of the electric field at the tip of the nanocone that allow the detection of a few molecules located near the tip end. The estimated Raman enhancement factor is about 106 with respect to a standard configuration. The present results open a good perspective for the development of an integrated Raman-AFM microscopy able to perform both topography and chemical mapping in label free condition with a spatial resolution comparable to the tip end.

Nanoplasmonic and Microfluidic Devices for Biological Sensing

In this chapter we report about recent advances on the development and application of 2D and 3D plasmonic nanostructures used for sensing of biological samples by Raman spectroscopy at unprecedented resolution of analysis. Besides, we explain how the integration of these nanodevices in a microfluidic apparatus can simplify the analysis of biological samples. In the first part we introduce and motivate the convenience of using nanoplasmonic enhancers and Raman spectroscopy for biological sensing, describing the phenomena and the current approaches to fabricate nanoplasmonic structures. In the second part, we explain how specific multi-element devices produce the optimal enhancement of the Raman scattering. We report cases where biological sensing of DNA was performed at few molecules level with nanometer spatial resolutions. Finally, we show an example of microfluidic device integrating plasmonic nanodevices to sort and drive biological samples, like living cells, towards the optical probe in order to obtain optimal conditions of analysis.

Aspirin wears smart

Low-dose aspirin is used worldwide for preventing thromboembolic disorders. Its use, however, is often associated with gastrointestinal bleeding, mostly due to direct irritation of the gastric mucosa. Here we provide evidence for a novel sublingual formulation of aspirin micronized and co-grinded with collagen proven to be as effective as oral standard formulation in inhibiting platelet aggregation but with attenuated gastric irritation. This represents a new option for better aspirin treatment in the prevention of myocardial infarction and stroke. Orally given low-dose aspirin has been used for decades due to its anti-inflammatory and antithrombotic properties. Although standard oral formulation of aspirin allows rapid and complete absorption from the GI tract, new formulations have been developed and marketed, (e.g. dry granules, effervescent solution, and chewable tablets) with the aim to achieve faster dissolution and faster absorption as well as to reduce direct aspirin-induced gastric lesions. However, the occurrence of gastrointestinal bleeding still remains a significant problem of chronic aspirin administration and, sometimes, limits the use of aspirin in primary prevention. On the other hand, co-administration of proton pump inhibitors is currently used to counteract aspirin-induced gastric lesions, thereby representing a pharmaco-economic issue in the area of health care sustainability. Recently, we developed and patented (N. 102015000079955) a new formulation of aspirin which leads to faster absorption and activity but devoid of direct gastrointestinal lesioning effect. In addition, this formulation allows sublingual administration of the drug which, by passing the liver metabolism, leads to

Superhydrophobicity, plasmonics and Raman spectroscopy for few/single molecule detection down to attomolar concentration

Few/single molecule detection is of great importance in fields including biomedicine, safety and eco-pollution in relation to rare and dangerous chemicals. Superhydrophobic surfaces incorporated with the nanoplasmonic structure enable this device to overcome the diffusion limit of molecules dissolved in water with the concentration down to 10 attomolar. In this paper demonstrated the fabrication of hydrophobic surfaces using optical lithography/reactive ion etching and its application to overcome the diffusion limit. Various experiments such as contact angle measurements, SEM, fluorescence, Raman and FTIR absorption spectroscopy were performed which indicate that utilizing this device it could be possible to perform the measurements for the sample with extremely low dilution. The major application of this novel family of devices would be the early detection of tumors or other important pathologies, with incredible advances in medicine.