An Original Method for Removing and Fixing the Pituitary Gland of Sexually Mature Rats for Creating High-Quality Permanent Histological Preparations

Autor: N. B. Hryntsova, Sumy State Univercity. , Rymskogo-Korsakova St., Sumy , Ukraine, A. M. Romanyuk
Rok vydání: 2021
Předmět:
Zdroj: Ukraïnsʹkij žurnal medicini, bìologìï ta sportu. 6:303-310
ISSN: 2522-4972
2415-3060
DOI: 10.26693/jmbs06.01.303
Popis: The issue of high-quality creating histological preparations is an urgent issue of histology and pathological anatomy, experimental medicine and biology. The pituitary gland belongs to the smallest in mass and size of the endocrine glands in both humans and animals, it is an endocrine gland and occupies one of the central places in the endocrine regulation of the body's vital activity. The study of the features of the removal of the pituitary gland from Turkish saddle of the sphenoid bone of the skull of rats remains an important aspect of modern morphology. The purpose of the work was to develop an original technique for improving the first (removal) and second (material fixation) stages of creating histological preparations of the rat pituitary gland for morphological, morphometric, and immunohistochemical medico-biological experimental studies. Material and methods. The development of the methodology was carried out on 200 white sexually mature rats of different sex weighing 250-300 g at the age of 7-8 months in accordance with the National and European bioethical standards. Results and discussion. Modification of the technique of extracting the pituitary gland of rats includes the introduction of the animal into thiopental anesthesia, decapitation, separation of the skin and muscular integument of the head, resection of the occipital bone of the skull, exposure and removal of the brain, identification of the pituitary gland, followed by removal of the pituitary gland with a single complex (block) together with the pituitary fossa Turkish saddle and fragments of the sphenoid bone adjacent to it. The gland was fixed with a reduced concentration of a 5% solution of buffered neutral formalin. After 15-18 hours from the beginning of fixation, the complex was temporarily removed from the fixator, and the pituitary gland was removed from the pituitary fossa of Turkish saddle using eye scalpel and eye forceps. The organ had a well-fixed and compacted capsule, which prevented unnecessary trauma and fragmentation of the organ during removal. The removed pituitary gland was again immersed in a 5% solution of neutral buffered formalin for 2-3 hours. After the end of the fixation period, the pituitary gland had a well-fixed structure and was subject to the following standard stages of making permanent histological specimens
Databáze: OpenAIRE