The Neuroanatomy and Experimental Histology Service provides the scientific community with its renowned professional experience in the fields of Neuroanatomy and Histology.
FREEZING AND CUTTING FROZEN SECTIONS OF HISTOLOGICAL MATERIAL
High-level Technology Service (STAN)
Description:
A tissue can be sliced into thin sections with a freezing microtome (30-60 microns) or in a cryostat (10-50 microns). The advantage of using the freezing microtome is that it performs quick cuts and, therefore, plenty of material can be processed daily. Moreover, in the cryostat the tissue does not undergo temperature fluctuations and also, the machine performs thinner cuts which are placed directly on the microscope slide for later analysis.
Methodology: The sample is placed on the microtome base and then a quick freezing is carried out with carbon dioxide. On a following stage, the suggested thickness is cut and the material is collected in corresponding solutions.
NEUROTOXICOLOGY
High-level Technology Service (STAN)
Description: Neuronal death detection with the use of neurohistological techniques for evaluation in the field of Neurotoxicology.
Methodology: The Silver Staining technique is of great value for neurotoxicological studies due to its high degree of sensitivity and fast temporal resolution of degenerative processes. Given its capacity to initiate in situ silver deposits, this technique allows to detect the degeneration of whole neurons and the loss of cellular components, constituting a highly useful tool for the study of nerve tissue and also accurate enough for the routinely observation of nerve cells, dendrites and axons.
PERFUSION OF ANIMALS
High-level Technology Service (STAN)
Description: The Neuroanatomy and Experimental Histology Laboratory offers the service of test animals´ perfusion. In this procedure, test animals are transcardially perfused and all the blood from the circulatory system is eliminated. Later, a cleansing and a fixative solutions are applied with the aim of optimizing the preservation of the structures and features of the tissue being studied.
IMMUNOHISTOCHEMISTRY ON ANIMAL TISSUE
High-level Technology Service (STAN)
Description: This service consists in the detection of cellular or tissue antigens by means of the antigen-antibody reaction. In order to visualize the location in which such reaction takes place, it is necessary to use a tracer or marker. The marking can be performed with fluorochromes (immunofluorescence techniques) or with colored substrates (immunoenzymatic techniques).
Methodology: Immunohistochemical techniques, widely used in neurohistological research, detect specific biomolecules (antigens) on tissue sections using specific antibodies (primary antibody) against the antigens detected. These primary antibodies are marked then with secondary antibodies that can be linked to a fluorochrome, which will subsequently enable the visualization with the fluorescent microscopy. Another alternative is to use biotinylated secondary antibodies that are later incubated in a solution with an avidin-peroxidase complex which reveals itself when exposed to a diaminobenzidine solution producing a brown colouring on the site of the antigen. This can be clearly visualized in the light microscope.
TRAINING AND COUNSELLING ON PREPARATION OF BIOLOGICAL SAMPLES FOR FLUORESCENCE MICROSCOPY
High-level Technology Service (STAN, in its Spanish acronym)
Description: The choice of the sample, the marking technique and the material to be used are crucial for the image documentation and analysis. Besides, the fact that new alternatives are being adopted with an unprecedented speed in the area of cellular marking, and the absence of this service at the national and regional level creates the need of training and counselling within this area. The service is aimed at researchers, health professionals and staff of companies that use or sell fluorescence microscopes.
Methodology: The counselling can be performed through onsite or distance learning options. Onsite counselling offers practice in preparation of histological and cytological (fixed or live) samples in immuno-marking and/or transfection techniques. Through distance learning, the counselling is online offering information about procedures in all cellular marking stages. Potentially, the service can be offered as a postgraduate course.
Responsable:
Dra. Soledad de Olmos
Phone Number: (54 – 0351) 4681461
e-mail: sdeolmos@immf.uncor.edu – neuroanatomia@immf.uncor.edu