The Lab on a Chip (LOC) family is wide and multifaced due to the large number of possible applications ranging from the biomedical to the agrifood and environmental fields. Due to their better performance in terms of speed, flexibility, automation and costs, LOCs present important advantages with respect to conventional methods. The huge practical potential of these platforms is confirmed by our interactions with several companies as active partners or sponsors in several national and international projects.
LOC for Medical Diagnostics
A major aim in clinical and proteomic research is the development of new tools for analysis and high throughput evaluation of markers of interest, able to allow a prompt and sensitive diagnosis of pathological and threatening conditions. In such a needing of fast response and low cost analysis, the development of new investigation instruments to replace standard, time-spending and complex laboratory tools, requires more and more features of miniaturization, easy-to-use and portability for point of care analysis.
At Nanotec we have developed different LOCs for diagnostic applications. In particular, merging microfluidic with biosensing modules, we have realized several biochips for diagnosis of human diseases. In particular biochips for cancer diagnostics (Prostate and Pancreatic cancer) have been developed. A LOC, consisting of microelectrode arrays and microfluidic components has been functionalized to hold antibodies or peptides in order to detect specific biomarkers. Recently we are working on the development of artificial receptors based on molecularly imprinted polymer (MIP) electrosynthetized on chip for small molecules detection. Then, biochips based on Electric Impedance Spectroscopy (EIS) have been used to perform studies/detection of several cells ranging from bacteria to human cells. In cooperation with Ekuberg pharma we developed a biosensing platform which allows a multiplexed and simultaneous detection of pathogens for a quick screening for lower genital tract infection. We also implemented a biochip for automatic tests to quantify invasive potential of cell lines, detect migratory activity associated with matrix degradation.
We also optimized microchannels functionalized with smart microgels for on chip release of fluorescent antibodies for cell staining and subsequent detection and counting. This system was produced by novel covalent immobilization of pH-responsive poly(methacrylic acid) microgels onto microfluidic channels. Afterwards, selected thiophene labeled monoclonal antibodies have been encapsulated and then released in presence of cells providing cell labeling.
Recently we produced an electrochromic“point-of-care” device for cystic fibrosis diagnosis, especially in ambulatory contests. Sodium chloride sweat promotes an oxide blue coloration proportional to the cation concentration in the sweat.
Part of the activities of this research line also aims to develop methods and devices for the physical isolation of different subclasses of Extracellular vesicles (EV) essentially based on physicochemical properties (size, charge, polarizability, rigidity etc.), rather than on molecular markers. EVs are a vastly heterogeneous class of cell-derived particles, spanning from 40 nm to 2 μm, which play a crucial role in several physiological and pathological conditions. Currently, the analysis of EVs is hampered by the lack of efficient and standardized technologies for their isolation and characterization. To this end, we are exploring combinations of hydrodynamic principles and various force fields, starting from theoretical simulations by finite elements methods followed by device fabrication by rapid prototyping and experimental testing. Furthermore, in association with Maastricht University, LOC approaches will be exploited to create an organ-on-chip platform to study degenerative and regenerative processes and develop more efficient therapies.
LOC for Food safety & Environmental Monitoring
Beyond diagnostics, the potential of lab-on-chip for the agricultural fields and environmental monitoring is also of huge interest, for example to monitor food safety along the whole production chain and to prevent environmental contamination. In this scenario, we have developed LOC devices and we have optimized three different applications in the control of foodstuffs. Specifically, we demonstrated an efficient detection of components involved in food allergies and in particular in the common celiac disease. For this study, an array of electrodes was functionalized with antibodies against gliadin, a prolamine of cereals and a gluten compound, responsible allergic reaction of people affected. Thanks to our biochip we were able to sensitively quantify the amount of gliadin on its specific antibody in samples derived from both liquid and solid matrix with a detection limit of 0.5 µg/ml. The second developed application deals with the detection of some of the most common bacteria involved in foodborne diseases such as Listeria monocytogenes and Staphylococcus aureus by using specific antibodies against antigens expressed on the bacterial cells surface in order to immobilize the whole bacteria on the electrodes. Foodborne diseases are responsible for high levels of morbidity and mortality in the general population, but particularly for high-risk groups, such as infants, children, elderly and immunocompromised people. The third application aims to detect the presence of mycotoxins contamination from food samples and in particular ochratoxin A (OTA), a toxic and potentially carcinogenic fungal toxin produced by Penicillium and Aspergillus species and found in a variety of food commodities, which elicit necrosis, hepatitis and immunosuppressive effect. A label free detection was achieved through the immobilization of a specific DNA aptamer onto the surface of gold trasducers with a dedicated functionalization.
In the environmental field, we have developed an EIS immunosensor for the sensitive, direct and label-free detection of cholera toxin with a limit of detection smaller than 10pM, a value thousands of times lower than the lethal dose.
Another environmental problem which needs to be monitored is the contamination of heavy metals. To respond to this need we have developed a chip based on anodic stripping voltammetry for the detection of copper. The metal of interest is preconcentrated on the working electrode during a deposition step, and oxidized from the electrode during the stripping step. The oxidation of metal is registered as a current peak at a specific potential.
LOC for pharmaceutical formulations
Another very active research field concerns “On-chip-chemistry” which opens huge possibilities to miniaturize and perform on chip complex synthetic processes to obtain chemicals or pharmaceuticals in continuous flow. An application developed in the past in the frame of the European Project “Radiochemistry On Chip” was a microfluidic platform able to synthesize radiopharmaceuticals for Positron Emission Tomography (PET) in a dose-on-demand setting within a few minutes. Compared with traditional systems, the microfluidic approach resulted to be faster (simplified synthetic route) with minimized reagent consumption, waste production and operator interactions with radioactivity. As shown in the figure below, glass microchannels have been designed to allow on-chip mixing (see Module 2 and 4 in the Figure), concentration (Module 1 in the Figure), solvent exchange (Module 3 in the Figure). Another application was a continuously operating microfluidic device for solvent removal and exchange for chemical syntheses by means of a supporting gas. It was demonstrated that acetonitrile was reduced from 50 wt% in the feed solution to 1 wt% in the final solution. On chip catalysis was also exploited to synthesize biaryls via Kumada Corriu cross-coupling in a glass microreactor with a silica supported nickel-based catalyst. The reaction time was dramatically decreased (four order of magnitude) and the yields increased threefold compared with batch reactions. In the frame of microfabrication, a microwave assisted-method for microchannel production was recently set to allow fast and safe production of microstructures for LOC applications. The research in this field is currently funded by a recent approved regional project aimed at producing a microfluidic apparatus for in-flow fast purification of radiopharmaceuticals.
Microfluidics for LOC applications
Transversal to all these activities is research on microfluidics which targets the manipulation and control of fluids (either samples or reagents) at the microscale and allows the design of Micro Total Analysis System (µ-TAS) for investigating a variety of physical, chemical and biochemical separation/purification processes. In particular, capillary motion in a confined environment allows to generate and control boundary effects, restricted diffusion and elastic turbulence, with several practical implication in LOC implementation. Studies of capillary dynamics in microchannels has been used to investigate the solid surface/liquid interactions and predict the behavior of aggressive solvents in teflon-coated polydimethylsiloxane (PDMS) microchannels. Analogue experiments have been performed to estimate the self-propelling properties of Ru4POM microcapsules able to induce elastic stress in aqueous solutions fed with H2O2. Droplet-microfluidics was exploited to study the properties of the natural hydrophobin protein HFBI as stabilizer of fluorous oil droplets in aqueous environment to prepare emulsified systems for biomedical applications. As regards µ-TAS, a membrane-based microfluidic system was produced to allow facile separation of red blood cells from whole blood to collect clean plasma. This technology sounds interesting in view of producing a Lab-on-a-chip for diagnostics that uses µL droplets of whole blood rather than out-of-chip produced plasma. Finally, innovative devices have been manufactured on a large variety of smart, flexible and biodegradable materials, such as biopolymers, paper and fabrics.
- M. S. Chiriacò, M. Bianco, F. Amato, E. Primiceri, F. Ferrara, V. Arima, G. Maruccio Fabrication of interconnected multilevel channels in a monolithic SU-8 structure using a LOR sacrificial layer Microelectronic Engineering 164, 1, 30-35 (2016) ISSN: 01679317DOI: 10.1016/j.mee.2016.07.006
- A. Zacheo, A. Quarta, A. Zizzari, A. G. Monteduro, G. Maruccio, V. Arima and G. Gigli One step preparation of quantum dot-embedded lipid nanovesicles by a microfluidic device RSC Adv., 5, 98576-98582 (2015) ISSN: 2046-2069; doi: 10.1039/c5ra18862h
- A .Zacheo, A. Zizzari, E. Perrone, L. Carbone, G. Giancane, L. Valli, R. Rinaldi and V. Arima Fast and safe microwave-assisted glass channel shaped microstructure fabrication Lab Chip 15, 2395–2399 (2015) ISSN: 1473-0197; doi: 10.1039/c4lc01419g
- V. Arima, P. Watts, and G. Pascali Microfluidics in Planar Microchannels: Synthesis of Chemical Compounds On-Chip Chapter 8 from “Lab-on-a-Chip Devices and Micro-Total Analysis Systems: A Practical Guide” Published: November 5, 2014 by Springer. Editor(s): by Jaime Castillo-León and Winnie E. Svendsen
- A .Zizzari, M. Bianco, R. Miglietta, L. L. del Mercato, M. Carraro, A. Sorarù, M. Bonchio, G. Gigli, R. Rinaldi, I. Viola, V. Arima, Catalytic oxygen production mediated by smart capsules to modulate elastic turbulence under a laminar flow regime Lab Chip, 14, 4391-4397 (2014) ISSN: 1473-0197; doi: 10.1039/c4lc00791c
- L. L. del Mercato, M. Carraro, A. Zizzari, M. Bianco, R. Miglietta, V. Arima, I. Viola, C. Nobile, A. Sorar, D. Vilona, G. Gigli, M. Bonchio, R. Rinaldi Catalytic Self-Propulsion of Supramolecular Capsules Powered by Polyoxometalate Cargos Chem. Eur. J. 20, 1 – 6 (2014) ISSN: 0947-6539; doi: 10.1002/chem.201403171
- V. De Matteis, A. Cannavale, L. Blasi, A. Quarta, G. Gigli “Chromogenic device for cystic fibrosis precocious diagnosis: A “point of care” tool for sweat test” Sensors and Actuators B: Chemical 225, 474–480 (2016) ISSN: 09254005; doi: 10.1016/j.snb.2015.11.080
- M. S. Chiriacò, F. de Feo, E. Primiceri, A. G. Monteduro, G. E. de Benedetto, A. Pennetta, R. Rinaldi and G. Maruccio, Portable gliadin-immunochip for contamination control on the food production chain Talanta 142, 57-63 (2015) ISSN: 00399140; doi: 10.1016/j.talanta.2015.04.040
Other selected publications
- I. Viola, N. Ghofraniha, A. Zacheo, V. Arima, C. Conti, G. Gigli Random laser emission from a paper-based device J. Mat. Chem. C 1, 8128-8133 (2013) ISSN: 2050-7526; doi: 10.1039/c3tc31860e
- V. Arima, G, Pascali, O. Lade, H. R. Kretschmer, I. Bernsdorf, V. Hammond, P. Watts, F. De Leonardis, M. D. Tarn, N. Pamme, B. Z. Cvetkovic, P. S. Dittrich, N. Vasovic, R. Duane, A. Jaksic, A. Zacheo, A. Zizzari, L. Marra, E. Perrone, P. A. Salvadori, R. Rinaldi Radiochemistry on chip: towards dose-on-demand synthesis of PET radiopharmaceuticals Lab on a chip 13, 2328-2336 (2013) ISSN: 1473-0197; doi: 10.1039/c3lc00055a
- R. Milani, E. Monogioudi, M. Baldrighi, Cavallo, V. Arima, L. Marra, A. Zizzari, R. Rinaldi, M- Linder, G. Resnati, P. Metrangolo Hydrophobin: fluorosurfactant-like properties without fluorine Soft Matter 9, 6505-6514 (2013) ISSN: 1744683X; doi: 10.1039/c3sm51262b
- N. Ghofraniha, I. Viola, A. Zacheo, A., I. Viola, A. Zacheo, V. Arima, G. Gigli, C. Conti Transition from nonresonant to resonant random lasers by the geometrical confinement of disorder Opt. Lett. 38, 5043-5046 (2013) ISSN: 0146-9592; doi: 10.1364/OL.38.005043
- B. Z. Cvetković, O. Lade, L. Marra, V. Arima, R. Rinaldi and P. S. Dittrich Nitrogen supported solvent evaporation using continuous-flow microfluidics RSC Adv. 2, 11117-11122 (2012) ISSN: 20462069; doi: 10.1039/c2ra21876c
- E .Primiceri, M. S. Chiriacò, E. D’Amone, E. Urso, R. E. Ionescu, A. Rizzello, M. Maffia, R. Cingolani, R. Rinaldi and G. Maruccio, Real-time monitoring of copper ions-induced cytotoxicity by EIS cell chips, Biosens. Bioelectron. 25, 2711-2716 (2010) ISSN: 09565663; doi: 10.1016/j.bios.2010.04.032
- M. S. Chiriacò, E. Primiceri, E. D’Amone, R. E. Ionescu, R. Rinaldi and G. Maruccio, EIS microfluidic chips for flow immunoassay and ultrasensitive cholera toxin detection, Lab on a Chip 11, 658-663 (2011). ISSN: 1473-0197; doi: 10.1039/c0lc00409j
- E. Primiceri, M. S. Chiriacò, F. Dioguardi, A. G. Monteduro, E. D’Amone, R. Rinaldi, G. Giannelli and G. Maruccio, Automatic transwell assay by an EIS cell chip to monitor cell migration, Lab on a Chip 11, 4081-4086 (2011). ISSN: 1473-0197; doi: 10.1039/c1lc20540d
- M. S. Chiriacò, E. Primiceri, A. Montanaro, F. de Feo, L. Leone, R. Rinaldi and G. Maruccio, On-chip screening for prostate cancer: an EIS microfluidic platform for contemporary detection of free and total PSA, Analyst 138, 5404-5410 (2013). ISSN: 0003-2654; doi: 10.1039/c3an00911d
- E. Primiceri, M. S. Chiriacò, R. Rinaldi and G. Maruccio, Cell chips as new tools for cell biology – results, perspectives and opportunities, Lab on a Chip 13, 3789-3802 (2013). ISSN: 1473-0197; doi: 10.1039/c3lc50550b
- Carati, M. S. Chiriaco, F. De Feo, G. Maruccio, M. Megha, A. Montanaro, E. Primiceri and A. Tinelli, WO2015015456-A1.
The object of the present invention is to provide an impedimetric biochip for the simultaneous diagnosis of gynecological pathologies related to C. albicans, S. agalactiae or C.trachomatis by using the vaginal fluid of the patient. This invention enables a significant reduction of examination times if compared with the present-day techniques. Another important feature of the present invention is its ability to provide a highly sensitive detection, accurate and specific for the described diseases.