This unit has facilities with a perimeter area of 3,200 m2 and it has large rooms to house the experimental animals according to their species characteristics. Among the equipment and facilities include:
9 rooms for large animals (pigs) maintenance with capacity for about 350 animals, for long periods of time and environmental, nutritional and microbiological conditions controlled.
2 rooms for the maintenance of small ruminants (sheep and goats) with capacity for about 60 animals over long periods of time and environmental, nutritional and microbiological conditions controlled.
Leica TCS-SP5 confocal microscope with especial features that allows studying interactions between cells/tissues and materials. Indeed, the experience of the research group in charge of this Unit makes this a unique service for the study of cells and tissues and the interactions between various materials and cell components as well as between implants/scaffolds and tissues of the recipient organism. For example, it makes it possible to characterize in detail the specific markers of particular cell populations for tissue engineering applications and in vitro tests of biocompatibility of new materials, allowing the process of cell colonization of surfaces to be explored and simultaneous analysis of the cells or subcellular structures and materials. The combined use of fluorescence and reflection makes it possible to study the tissue-implant interface, with up to 8 fluorophores, at wavelengths from 405 to 633 nm allowing histological studies of a wide range of tissue types which would be restricted at certain fixed wavelengths due to autofluorescence phenomena.
Leica TCS-SP5 confocal microscope with:
DMI 6000 inverted microscope with 4 objectives:
10X (dry); 20X; 40X and 63X(for immersion).
Confocal module:
•• 3 channels for spectral detection
•• AOBS (Acousto Optical Beam Splitter)
•• Resonant scanning system.
4 lasers:
•• Argon laser (excitation at 458, 477, 488, 496 and 514 nm)
•• He/Ne laser (excitation at 633 nm)
•• DPSS (diode-pumped solid-state) laser (excitation at 561 nm)
•• UV laser (excitation at 405 nm)
Incubation chamber on a motorized stage.
Workstation with 4 types of software for data acquisition and processing:
•• 3D imaging
•• Colocalization analysis
•• FRAP (Fluorescence Recovery After Photobleaching)
•• FLIP (Fluorescene Loss In Photobleaching)
•• FRET (Fluorescence Resonance Energy Transfer)
ToF-SIMS 5 Time-of-flight secondary-ion mass spectrometry system (ION TOF ).
•• Sensitivity < 100 ppm.
•• Lateral resolution ~ 100 nm (extension of surface area analyzed).
•• Depth resolution ~ 1 nm (thickness of surface analyzed).
•• Identification of molecules up to 10000 amu.
K-Alpha X-ray photoelectron spectroscopy (XPS) system (Thermo):
•• Lateral resolution ~ 30-400 μm (extension of surface area analyzed).
•• Depth resolution ~ 10 nm (thickness of surface analyzed).
•• Depth profiles.
UVISEL ellipsometer (Horiba JovinYvon), range 190-2100 nm, based on phase modulation with cell for liquid media:
•• Measurement of thicknesses from 0.1 nm to 30 μm.
•• Measurement of thickness of various superimposed layers.
•• Simultaneous measurement of the proportion of various different materials present on a surface.
•• Estimation of the porosity as percentage of pores vs. deposited material.
•• Measurement of roughness of coating between 2.5 and 15 nm.
Temporarily OUT OF ORDERTAM III isothermal nanocalorimeter system including options for titration and perfusion. Also solution quasi-adiabatic calorimetry is available. (TA Instruments). Thermostatic bath: •• Working temperatures between 15 and 150 ºC •• Temperature scan < 2 ºC/h •• Stability: /- 10 μK on short timescales and /- 100 μK drift over 24 hours Calorimetric response: •• Precision better than 100 nW •• Reproducibility better than 1% •• Stability: < 10 nW on short timescales and < 40 nW drift over 24 hours Solution: •• Maximum temperature 80ºC •• Quasi-adiabatic mode of operation
T1 and T2 Nuclear Magnetic Resonance Relaxometry:
Stelar SmarTRACER (Italy) + Bruker (Germany) 2 T electromagnet. Fast Field Cycling designed to measure longitudinal nuclear magnetic relaxation as a function of the magnetic field intensity. Measurements of the longitudinal (T1) and transverse (T2) time constants as a function of the Larmor frequency.
•• Measurement range: continuous measurement from 10 kHz (almost null field) to 10 MHz (0.25T) to obtain T1.
•• Measurement range: measurement from 10 MHz (0.25T) to 80MHz (1.9T) at desired intervals to obtain T1 and T2.
•• Inhomogeneity lower than 150 PPM.
•• Main pulse sequences implemented with the possibility of modifying parameters to the design and programming of new sequences.
•• Temperature control from -120°C to +140°C with accuracy and stability of 0.1°C.
Alternating Gradient Magnetometer:
MicroMag M2900-4 AGM (Princeton Measurements Corporation, USA) for the magnetic and mechanical characterization of nanoparticles in various different media:
•• Magnetic Moment Range: 1nA.m² to 5mA.m² full scale (1μemu to 5emu).
•• Resolution: 0,005% of full scale with 60% overrange capability.
•• Speed of measurement: 100 ms/point.
•• High sensitivity: 10 pA.m² (10 nemu) of standard deviation at room temperature and with 1 second of averaging time.
•• High performance with samples of very small magnetization and dimensions (few nanometers).
•• The AGM can accommodate a large range of samples with widely different properties.
Specifications:
›› Three heads for injection of different materials:
•• Polymers
•• Ceramics
•• Living cells
›› Allowed viscosities ranging from 1 to 106 centipoises.
›› System for temperature and humidity control to enable the printing conditions to be adjusted. It is possible to create a microenvironment appropriate for each mixture of materials and proteins and to achieve high rates of cell viability.
›› Controlled by Software .
›› Visual monitoring system based on a digital camera, firewire cameras and high magnification lenses, with detection and measurement of colormetric/monochromatic variations.
›› Mapping system using a laser sensor for control of normal and extended working distances. It includes Target, Grid and Path mapping.
›› High-precision dispensing pump for dynamic control of the flow rate.
›› Position control system:
•• X/Y/Z accuracy: ±10 μm
•• X/Y reproducibility: ±2 μm
•• X/Y resolution: 0.5 μm
•• X/Y velocity: 304 mm/s (12”/s)
•• X/Y displacement: 304X 152 mm (12”X6”)
•• Z displacement: 101 mm (4”)
In addition, it should be noted that the system is sited in a clean room with a controlled environment and supply of the gases required for these tests.
