Histološka analiza organa potočne pastrve (Salmo trutta Linnaeus, 1758) fiksiranih različitim fiksativima

master's thesis

• Title (english): Histological analysis internal organs of brown trout (Salmo trutta Linnaeus, 1758) fixed by various fixators
• Author: Ljerka Armano
• Mentor: Romana Gračan (mentor)
• Committee member: Željka Vidaković Cifrek (predsjednik povjerenstva)
• Committee member: Duje Lisičić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2019, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Fiksiranje tkiva važan je korak u histološkoj obradi preparata. Svrha fiksacije je očuvanje stanica i međustaničnih struktura kako bi što više nalikovali živom materijalu. Istraživanje je provedeno na uzorcima tkiva probavnog, dišnog i cirkulacijskog sustava potočne pastrve (Salmo trutta L.). Uspoređivala se morfološka struktura glavnih unutrašnjih organa potočne pastrve fiksiranih s dva različita fiksativa: neutralni puferirani formaldehid (PNF) koji daje kvalitetne histološke uzorke, ali je toksičan i kancerogen te zamjenskog fiksativa FineFix-a (FF) smanjene toksičnosti baziranog na etanolu. Za što bolju usporedbu kvalitete obojenosti tkiva, korištena je hemalaun eozinska (HE) tehnika te histokemijska metoda bojenja perjodne kiseline i Schiffovog reagensa te alcijanskog modrila s obzirom na dva istraživana fiksativa. Rezultati ukazuju da je 10% PNF i dalje najbolji izbor pri standardnoj fiksaciji za histološke i histokemijske analize. Kao najveća razlika između dva upotrebljena fiksativa ističe se kvaliteta fiksacije vezivnog tkiva u proučavanim organima, koja je bolja u tkivima fiksiranim s PNF, u usporedbi s FF, dok se FF istaknuo kao bolji promatranjem HE preparata probavnog sustava. Stoga određena osjetljiva tkiva poput probavnih organa zahtijevaju modificiranje standardnih postupaka bojenja, a naročito HE metode, kako bi se osigurao bolji afinitet tkiva za boju, a time i bolje prepoznavanje morfoloških struktura u tkivu.

Abstract (english)

Tissue fixation is a very important step when preparing permanent histological samples. The purpose of fixation is to preserve cells and extracellular structures to resemble living material as much as possible. We performed study on tissue samples of the digestive, respiratory and circulatory system of brown trout (Salmo trutta) to compare the histomorphological structures of major internal organs of brown trout fixed with two different fixatives: neutral buffered formaldehyde (PNF), which produces high-quality histological samples, but is toxic and carcinogen and FineFix (FF) ethanol-based substitution for PNF with reduced toxicity. For proper comparison of two studied fixatives, we performed tissue staining with routine hemalaun eosin (HE) technique and routine histochemical staining method for Periodic Acid - Schiff's reaction combined with alcian blue. Our results show that 10% PNF is a better choice of fixation for histological and histochemical analyses. The major difference between the two fixatives is the quality of connective tissue in the studied organs, which is better in PNF-fixed tissues compared to FF, while FF stands out as better when staining digestive system organs with HE. Therefore, certain sensitive tissues, such as the digestive organs, require modification of standard staining procedures, and in particular the HE method, to provide better tissue affinity for color and thus better recognition of tissue morphological structures.

• Keywords: fiksativi; formalin; formaldehid; FineFIX; fiksacija
• Keywords (english): fixative; formalin; formaldehyde; FineFIX; fixation
• Language: croatian
• URN:NBN: urn:nbn:hr:217:307645
• Study programme: Title: Biology; specializations in: Ecology; Course: Ecology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra biologije (magistar/magistra biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2019-11-20 08:05:27