centro biotecnologie avanzate

LABORATORY OF:Nanotechnologies / NANOMED
CONTACT PERSON:Prof Ugo Valbusa (University of Genova)
Phone +39 010 5737561 E-mail: valbusa@fisica.unige.it

Description of Laboratory and Expertise:

NANOMED labs aim to develop advanced nanotechnology promoting new knowledge in the fields of genomics, post-genomics and biomedicine in general.
Laboratories group multidisciplinary expertise in molecular biology, biochemistry, genetics and physics and include companies operating in the market of biotechnology and advanced electronics.

Abstract of Activities:

Low-cost and high-throughput gene expression profiling methods;
highly reproducible methods to analyze specific antigen expression patterns and protein interactions in cells and sera;
DNA genotyping and haplotyping by detecting individual probes on single strands;
high-throughput measurements of membrane potentials and ion channels activity on large scale or at single-events level;
mass selection and characterization of macromolecules by nano-molecular sieves.

Detailed Research Activities:

Nanopore Array (NPA)

The Human Genome project allowed the identification of new genes and Expressed Sequence Tags (EST) that will complete the decoding of the human genome. In the mean time, new technologies for the genome analysis have been developed, i.e. microarray technology, able to analyze ten thousands genes in a single experiment. In the 1990s, it was proposed that it might be possible to use biological nanopores (transmembrane proteins) as sensors for DNA.
The idea of the basic experiment is simple: DNA is a highly charged molecule, so it can be driven through the nanopore in a linear head-to-tail fashion by an electric field. When the DNA enters the nanopore, the ionic current is reduced because part of the liquid volume that carries the ionic current is occupied by the DNA. This might serve as a device to read off the sequence of DNA in an ultrafast way. More recently, non-biological, robust solid-state nanopores have opened up an even wider range of new research areas thanks to the fact that they could be ideal tools for a high-throughput platform for the molecular diagnosis of tumors and other genetic diseases. The NPA project aims at realizing a “nanopore array” device for low cost and high throughput gene expression profiling, exploiting the possibility offered by the Focused Ion Beam apparatus available in the lab to realize stable arrays of pores with controlled dimension on insulating substrates.

Genotype-Haplotype Microscopy

Atomic Force Microscopy (AFM) represents a powerful tool for morphological and structural characterization of single DNA molecules under different environments. It has been shown that high-resolution AFM imaging can be efficiently used for haplotyping DNA molecules, a results representing a significant advance over conventional approaches and which could facilitate the use of single-nucleotide polymorphisms (SNPs) for association and linkage studies of inherited diseases and genetic risks. Advances on this topic regard a proper understanding of: i) DNA molecules adsorption on solid surfaces, ii) the highest attainable AFM resolution with conventional imaging modes; iii) SNPs labeling strategies.

The GHM project focuses on the development of experimental protocols and instrumentation for AFM molecular haplotyping, with particular interest for high resolution AFM imaging of DNA molecules adsorbed on solid substrates and the design and development of a new AFM to achieve molecular resolution on DNA.

Nano-Patch-Clamp

The Nano-Patch-Clamp (NPC) project is aimed at building a high-throughput system dedicated to measures of ionic current in eukariotic cells. The analysis of ion-channel genes, and the screening of pharmaceutical compounds that affect specific ion channels, both require much higher-throughput assays of ion-channel activity than the traditional patch-clamp technique. Emerging chip-based technologies, especially the planar patch-clamp technology, are beginning to make large-scale screens of genes and compounds possible. In this context the know-how and the of technology available in the NanoMed research group (Focused Ion Beam, FIB) and the capabilities acquired on soft lithography techniques are the key factors for the development of a novel system of planar patch clamp.

Nano-Molecular-Sieves

Efficient methods to separate and analyze the different components of a mixture of biological macromolecules are of paramount importance, both for research and biomedical applications. To meet this need, a concerted effort is being made to develop miniaturized bioanalytical devices, using the technology of microfabrication.
These devices aim at superseeding the conventional methods based on gel electrophoresis, which are time consuming and difficult to automate. Using a gel to sieve the DNA complicates the procedure: a new matrix must be made each time, and recovery of the DNA is awkward, since the gel must either be sliced or blotted. Often, the reproducibility of the method is poor.
Using micro and nanofabrication techniques to implement molecular sieves gives several advantages: the spatial regularity of the sieve, the possibility of tuning the obstacle pattern on the size of the molecules to be separated, and of integrating the structure in a “lab-on-chip” system.

Applications and Developments:

Nano-Pore-Array (NPA) technology.
Genotype-Haplotype Microscopy (GHM).
Nano-Patch-Clamp (NPC) technology.
Nano-Molecular-Sieves (NMS) technology.

Managed core facilities:

Ultra High Resolution Field Emission Scanning Electron Microscopy (UHR-FE-SEM) and Focused Ion Beam (FIB), model CrossBeam® 1540XB by ZeissSEM Resolution 1.1nm, FIB Resolution 7nm.
Atomic Force Microscope Explorer by Thermomicroscopes - Veeco. (Explorer head, 100um tripod & 2um tube scanners, ECU+ control unit).Scanning-probe AFM suitable for morphological, mechanical and tribological characterization of samples down to the nanometer scale. It operates in a glove-box chamber under controlled atmosphere and humidity or in liquid environment. Operating modes: C-AFM, LFM-AFM, NC-AFM, Phase Imaging, Force - Volume.
Atomic Force Microscope Dimension 3100 by Digital Instruments - Veeco.(Dimension 3100 platform, XYZ Hybrid Head, Nanoscope IVa control unit).Scanning-probe AFM for single molecule characterization and manipulation. It is placed in a glove-box chamber for controlled atmosphere and humidity; it operates also in liquid environment. Operating modes: C-AFM, LFM-AFM, NC-AFM.
Fourier Transform Infrared Spectrometer (FTIR-THERMO) for optical characterization of samples in the infrared spectral range

Ongoing collaborations:

Biopolo, Milano;

Biubi s.r.l., Genova;

Diatheva s.r.l., Fano;

IFOM-FIRC Institute of Molecular Oncology, Milano;

IRCCS G. Gaslini, Genova;

IST National Institute for Cancer Research, Genova;

Laboratory Advanced Microscopy Bioimaging Spetroscopy, Genova;

MDM Materials and Devices for Microelectronics, Agrate Brianza;

Technobiochip Scarl, Latina;

UNIGE University of Genova, Genova;

UNIURB University of Urbino, Urbino

Most recent and significant publications

Mussi V., Granone F., Marolo T., Montereali R.M., Boragno C., Buatier de Mongeot F., Valbusa U. Surface nanostructuring and optical activation of Lithium fluoride crystals by ion beam irradiation. Appl Phys Lett. 2006 88, 103116.

Molle A., Buatier de Mongeot F., Molinari A., Boragno C., Valbusa U. Interfacial dynamics of the Rhomboidal pyramid pattern on the ion-eroded Cu (110) surface. Phys Rev. 2006 B 73, 155418.

Bruschi L., Fois G., Pontarollo A., Mistura G., Buatier de Mongeot F., Torre B., Boragno C., Buzio R., Valbusa U. Structural depinning of Ne monolayers on Pb at T<6.5K. Phys Rev Lett. 2006 96, 216101.

Buzio R., Boragno C., Valbusa U. Friction laws for lubricated nanocontacts. J Chem Phys. 2006 125, 094708.

Buzio R., Toma A., Chincarini A., Buatier de Mongeot F., Boragno C., Valbusa U. Atomic force microscopy and X-ray photoelectron spectroscopy characterization of low-energy ion sputtered mica. Surface science 2007 601, 2735-2739.

Felici R., Jeutter N.M., Mussi V., Buatier de Mongeot F., Boragno C., Valbusa U., Toma A., Wei Y., Rau C., Robinson I.K. In-situ study of the dewetting behavior of Ni-films on oxidized Si (001) by GISAXS. Surface science 2007 - in press.

Buzio R., Bosca A., krol S., Marchetto D., Valeri S., Valbusa U. Deformation and adhesion of elastomeric poly(dimethylsilonaxe) colloidal AFM probes. Langmuir 2007 23, 9293.

Moroni R., Buzio R., Chincarini A., Valbusa U., Buatier de Mongeot F., Bogani L., Caneschi A., Sessoli R., Cavigli L., Gurioli M. Optically Addressable Single-molecule-magnet behaviour of vacuum-sprayed ultrathin films. Journal Material Chem - in press.

Szymonski M., Goryl M. Kolodziej J.J., Buatier de Mongeot F. Metal nanostructures assembled at semiconductor surfaces studied with high resolution scanning probes. Nanotechnology 2007 18, ++044016.

Buatier de Mongeot F., Toma A., Molle A., Lizzit S., Petaccia L., Baraldi A. Self-organised synthesis of Rh nanostructures with tunable chemical reactivity. Nanoscale Res Lett. 2007 2, 251.

Further information is available at: http://www.nanomed.unige.it official site of Nanomed labs

ABC-Based grants, contracts, services

Agency/Company	Starting date	Expiring date	Title
industrie De Nora	16/11/07	15/11/08	Analyses on electrodes of industrial interest, aiming at identifying their chemical and physical properties