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INFRA-

STRUCTURE

MORPHOLOGY

IRET is equipped with a laboratory for the preparation of tissue sections, histological, immunohistochemical and immunofluorescence staining, with specific skills in multiple staining techniques.

Key instruments: Leica CM1950 motorized cryostat

CELL BIOLOGY

IRET has two separate rooms specifically for cell culture. In addition to cell lines, the laboratory uses primary cultures of neurons and glial cells, neural stem cells from fetal and adult brains, and animal embryonic stem cells. It uses induced pluripotent stem cells for various purposes, including the development of 3D disease models, and offers a vast range of models for in vitro disease studies, co-culture systems, cultures prepared on biomaterials, etc.

Key instruments: ICN Biomedicals Gelaire laminar flow cabinet, model TC48; POLARIS laminar flow hood, model 72; THERMO Forma incubator, model 1293; Sigma BINO stereo microscope; Olympus inverted microscopes, models IX70 and IX81.

ADVANCED MICROSCOPY

IRET is equipped with the most modern microscope imaging technologies, single-point and multiple laser confocal microscopy, usable also on living cells.

Key instruments: Nikon AX1 confocal laser scanning microscope, also configured for time-lapse experiments on multiple points, FRAP, FRET and calcium imaging modules; transmitted light and fluorescence microscopes, with motorized table and the following digital cameras: Nikon Eclipse E600, Nikon Eclipse Ni-E, Olympus AX70 TRF PROVIS, Nikon MICROPHOT-FXA.

2, 3, AND 4D IMAGE ANALYSIS

IRET is equipped with software for image analysis and quantification (2, 3 and 4D) and a cell-based high-content screening (HCS) analyzer for the automatic interpretation of cell culture plates.

Technological resources: 2, 3 and 4D image analysis software (Image ProPlus, Olympus Cell^P, Nikon NIS, Bitplane Imaris, Thermo Scientific CellInsight NXT); Thermo Scientific Cell Insight NXT HCS Platform; Thermo Scientific HCS Studio software.

PROTEOMICS

IRET is equipped with instruments for the analysis of proteins using methods based on immunochemical reactions (Western Blot; ELISA; xMAP technology), applicable to the quantification of proteins in different types of biological fluids and extracts. It participates in clinical studies in accordance with EMA/FDA specifications.

Key instruments: LUMINEX 200 equipped with xPONENT 3.1 software; MAGPIX equipped with xPONENT 4.2 and Milliplex Analyst 3.5 softwares; Adaltis Personal Lab automated ELISA analyzer and conventional spectrophotometer; Bio-Rad ChemiDoc MP imaging system.

MOLECULAR BIOLOGY

IRET offers equipment and skills for the mRNA analysis via RT-PCR and RT-PCR array of specific genes of interest or pathway-specific gene series, including on areas and cells microdissected from tissue sections or cell cultures. It produces tools for the manipulation of gene expression and is equipped with automated equipment for the extraction and assessment of nucleic acid quality.

Key instruments: Bio-Rad RT-PCR CFX-96; Thermo Scientific Nanodrop 2000;  QIAcube Connect System PRV-1 extractor (Qiagen); Qiaxcel Advanced analyzer (Qiagen); Nikon Eclipse TE2000-S+mmi Cellcut microdissector.

NGF RESEARCH CENTER

IRET is equipped with a laboratory specifically for the extraction of proteins from biological matrices (ÄKTA™ start protein purification system, Frac30 fraction collector, UNICORN START 1.1 software), with a particular focus on the extraction of nerve growth factor (NGF) according to the protocol developed by Dr. Luigi Aloe and Professor Rita Levi-Montalcini, a process which provides protein for in vitro and in vivo studies on the therapeutic efficacy of NGF.

ANIMAL FACILITY

IRET has a large facility for preclinical testing. The facility consists of four rooms for the housing of rats and mice (including transgenic animals), including metabolic cages and rooms for performing microsurgery and stereotaxic surgery on the brain and spinal cord. The facility has its own animal welfare body (OPBA, Organismo Preposto al Benessere Animale), for the lodging of applications for the authorization of animal testing with the Italian Ministry of Health.

Key instruments: operating microscope (Wild Heerbrugg, Wild M651); table for stereotaxic surgery on the spinal cord (Kopf) and brain (UGO BASILE); gas anesthesia stations (UGO BASILE); surgical instruments, osmotic minipump, peristaltic pump with UGO BASILE controller, LEICA Impact One.

ANIMAL BEHAVIOR

IRET is equipped with facilities for the study of complex behaviors via automated systems for the video recording and analysis of the sensory systems, locomotor function, and cognitive functions such as learning and memory, problem solving and attention.

Technological resources: dedicated testing areas and videotracking and analysis systems; Novel Object Recognition (NOR), Samsung; Contextual Fear Conditioning (CFC), UGO BASILE, model 46003; Y-Maze, Samsung; Water-maze, PanLab, Smart 2.5.16 model; Rota-rod, PanLab s.l., model LE8500; CatWalk, version 10.5, Noldus Information Technology.

GLP FACILITY

Through the social enterprise TransMed Research srl (of which IRET is a shareholder), IRET carries out in vivo and in vitro toxicity studies certified in accordance with the Good Laboratory Practice quality system (https://transmed-research.com).