The steep increase in the microscopy techniques, as well as their complexity in terms of sample preparation and analysis, calls for a centralisation of resources, in order to users to have one reference center where several expertise are available. PICO (Platform for Imaging at Cavalieri Ottolenghi) is located at the Neuroscience Institute Cavalieri Ottolenghi.
The Facility is equipped with excellent quality light microscopes , in particular, two confocal microscopes (Leica SP5 and Nikon) a Nikon ViCo system, and a high throughput Zeiss Axioscan image acquisition system. Moreover, a two-photon microscope Nikon (A1MP) and Light Sheet microscope (LaVision) have been recently installed .
The facility is also equipped for electron microscopy preparation and imaging, with one TEM available.
There are also several imaging systems equipped for high-resolution morphometric investigations, densitometry quantitative autoradiography, as well as workstations for image processing and statistical analysis. Two Neurolucida systems are included in the microscopy facility. Sliding or rotational microtomes, 3 vibratomes and 4 cryostats are available for neuro-histological studies.
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MOLECULAR AND CELLULAR NEUROBIOLOGY
A confocal microscope and numerous operating and research quality microscopes are available. An electron microscope is also accessible from the Dept. of Cell Biology, in the adjacent San Luigi Gonzaga Hospital. Several computer imaging systems with microscope and digital cameras are available for morphometry, autoradiographic quantitative densitometry, image processing, and statistical analysis. Neurohistology facilities offer sliding microtomes, rotary microtomes, vibratomes, and cryostats. The animal facility includes rooms for animal breeding, housing and behavioural testing and a room dedicated to rodent surgery.
Cellular and molecular biology
NICO has excellent facilities for research in molecular and cellular biology, which offer dedicated equipment and expertise for tissue culturing and molecular biology experiments.
The cell biology facility includes cell counting, cell freezing, cell plating for tissue culture experiments, and cell transfection. Inverted microscopes and time-lapse imaging system for living cells are available for in vitro and ex-vivo culture analyses (primary cultures, tissue explants, organotypic cultures, neurophere assay).
In addition, NICO offers expertise and services related to molecular biological techniques, such as DNA, RNA and microRNA preparation and analysis. The molecular biology facility includes a semi-automated nucleic acid purification platform, three Real-Time PCR machines, an electroporator for bacteria or ES as well as other standard laboratory equipment for extraction and analysis of DNA, RNA and proteins.
The neurophysiology laboratory is equipped for the preparation of microslices of nervous tissue, which are oxygenated and maintained in vitro for several hours.
Two experimental set-ups are available for patch-clamp recording of membrane potential or of ionic currents in single neurons in slices. They are equipped for electrical stimulation of axons and for application of pharmacological substances. Extracellular recordings are also available for studies of synaptic plasticity.
Behavioural tests facility
The laboratory is equipped with mazes and infrared camera for behavioural analysis of locomotor activity, anxiety, depressive-like behaviour and memory. A computerized video-tracking system (Ethovision XT video track system) is also available to analyze digitized images of behavioural tests.
CLINICAL NEUROBIOLOGI LABORATORY (CNL)
The CNL offers internal (University Hospital San Luigi Gonzaga) and external (10 neurology departments in the region) diagnostic and advisory service for the diagnosis of multiple sclerosis. Among the offered diagnostic tests there are the cerebrospinal fluid cytochemical examination, immunoisoelectrofocusing for searching oligoclonal bands, and different assays for viruses nucleic acid research. In addition, the laboratory provides a diagnostic service for paraneoplastic anti-neuronal antibodies.
Currently, the CNL is one of the few laboratories in Italy able to provide a diagnostic service for the detection of NMO-IgG and anti-AQP4 antibodies.
Finally, the CNL provides several services for biological monitoring of patients with multiple sclerosis treated with different drugs. In this regard, the laboratory runs a service in Italy and in Europe for serologic titration of antibodies against interferon-beta (by using three different methodologies) and natalizumab (Tysabri) possibly developed by patients treated with these drugs. Also, the laboratory provides a service for the evaluation of interferon-beta biological activity through the gene expression measurement of a specific interferon-induced protein (i.e. MxA).