Bioprinting

Bioprinting is a technology that creates living cellular constructs through the layer-by-layer, 3D deposition of biological materials, consisting in cells and extracellular matrix components (the bio-ink). It finds application in the development of biomimetic tissue models for in vitro studies or for the production of functional and transplantable tissue substitutes in Tissue Engineering. In particular, the Centre is developing a bioprinting system based on the microfluidic extrusion of cell-laden hydrogel fibres. Different hydrogels with tuneable chemical and mechanical properties are used as supporting extracellular matrix to recreate a 3D, instructive environment for cultured cells.

Flow cytometry

The flow cytometry laboratory  is equipped with two laser-based instruments which main purpose is to characterize and isolate a variety of cell population both of human and mouse origin. The technology of these instruments allows simultaneous multi-parametric analysis of thousands of cells for second for rapid analysis of complex cell populations. This is achieved by the use of different labelled antibodies or dyes to target specific molecules express at the surface or in the cytoplasm of the cell, giving a screening able to describe in detail a complex sample or to identify low frequency subset of cells.
Through these instruments, a variety of fluorochromes attached to different antibodies can screen the molecular components of a cell first to characterize it (flow cytometer) and then to specifically select for isolation (cell sorter).
The BD LSRFortessa cell analyzer is a 5 laser cytometer (488nm, UV 355nm, V 405nm, 561nm and 640nm) and 18 fluorescence detectors running a BD FACSDiva software on XP.

The characteristics of the cell analyzer are:

• to identify cells using antibodies for specific surface and intracellular markers;
• to discriminate transfetted cells using specific probes (i.e. GFP, RFP etc);
• to study cell proliferation (CFSE…) apoptosis and/or cell death (Annexin-V, 7-AAD). The BD FACSAriaIII is a 4 laser cell sorter (488nm, Near UV 375nm, 561 nm and 640nm) and 10 fluorescence detectors running a BD FACSDiva software on XP.

The details of the cell sorter are:

• up to 4 population cell sorting;
• single cell isolation;
• it can sort into tubes (1,5ml, 5ml, 15ml), multiwell plates (6, 24, 48, 96 and 384 wells), chamber slides (2 or 8 wells) and standard or frosted-end slides.

Microscopy

Imaging includes both commercial and state-of-the-art facilities, aimed at providing a complete panorama of techniques to tackle biomedical research activities. These include:

• Single and two-photon laser-scanning confocal microscopy.
• Spinningdisk confocal microscopy.
• Non-linear microscopy for CARS and SRS imaging.
• nanoIR microscopy.
• nanoRaman microscopy.
• Structured light microscopy.
• Optogenetics.

Micro and nanofluidic

The main purpose of the lab is to design micro and nanofluidics systems for biomedical applications. We are presently working on the development of a blood vessel model on chip and on the design of nanopore based sensors for single molecule protein and peptide analysis. In addition, we are involved in some fundamental science research lines on cavitation, microswimmers and multiphase flows at microscale.
The lab is equipped with a micro-PIV set-up for measurement of the flow velocity field at microscale (pulsed laser, inverted microscope, dual-frame camera), a high-speed camera and a set of pumps for manipulating small volume of fluids. The lab runs also in numerical simulations ranging from atomistic model to continuum fluid dynamics.

Cell culture

A cell culture facility supports the research of the two main projects by providing the appropriate equipment for the mammalian cell culture work. The purpose of the facility is to provide a common platform to carry out cell-based experiments and to maintain and to store experimental cell lines. A flow laminar hood with a stereoscope dedicated to cell reprogramming is dedicated to mouse and human pluripotent stem cells.