HISTOLOŠKE I HISTOKEMIJSKE TEHNIKE IDENTIFIKACIJE MASTOCITA KOD RIBA HISTOLOŠKE I HISTOKEMIJSKE TEHNIKE IDENTIFIKACIJE MASTOCITA KOD RIBA

master's thesis

• Title (english): HISTOLOGICAL AND HISTOCHEMICAL IDENTIFICATION OF MAST CELLS IN FISH
• Author: Mladenka Vukšić
• Mentor: Snježana Kužir (mentor)
• Mentor: Emil Gjurčević (mentor)
• Committee member: Krešimir Matanović (predsjednik povjerenstva)
• Committee member: Emil Gjurčević (član povjerenstva)
• Committee member: Snježana Kužir (član povjerenstva)
• Committee member: Lucija Bastiančić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Veterinary Medicine; Zagreb
• Defense date and country: 2024-12, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Veterinary Medicine; Basic and Preclinical Veterinary Sciences

Abstract

Mastociti riba su poznate, ali teže dokazive stanice. To je kroz povijest njihovog istraživanja dovelo
do raznih nedoumica, kao i do naziva eozinofilno granulirane stanice (engl. eosinophilic granular
cells/eosinophilic granule cells) kojim su pojedini autori označavali ove stanice. U recentnim istraživanjima
ovaj je naziv napušten. Svojstvo metakromazije granula mastocita riba često nije izraženo. Prema
dostupnoj literaturi, uzrok tomu može biti sastav njihovih granula ili njihova moguća topivost nakon
primjene vodenih fiksativa. Tinktorijalna svojstva granula mastocita pokazuju velike razlike među vrstama,
među dobnim skupinama unutar iste vrste i ovisno o zdravstvenom stanju.
Cilj ovog istraživanja bio je prikazati prikladnost određenih fiksativa u kombinaciji s bojenjima pri
vizualizaciji mastocita kod štuke (Esox lucius). Za potrebe izrade diplomskog rada korišteni su parafinski
blokovi s uzorcima organa od ukupno tri štuke podrijetlom iz rijeke Mrežnice. Iz svakog pojedinog
primjerka izdvojeni uzorci probavnih organa fiksirani su u četiri fiksativa: 10 % neutralni formalin, Bouinov fiksativ, 100 % etanol i Carnoy-ev fiksativ. Obojeni su s histološkim i histokemijskim metodama
(hematoksilin eozin, toluidinsko plavilo, alciansko plavilo-periodično bojenje kiselinom po Schiffu,
alciansko plavilo-safranin i Verhoeff -Van Giesen).
Korištenim fiksativima i toluidinskim plavilom nije dobivena metakromazija granula mastocita kod
štuke. Korištenjem 10 %-tnog neutralnog formalina i bojenjem toluidinskim plavilom moguće je dobiti
jasnu vizualizaciju mastocita s ortokromatski obojenim granulama. Etanol se pokazao kao neprikladan
fiksativ za crijeva, dok su Carnoy-ev i Bouin-ov fiksativ prikladni za dobivanje izražene eozinofilije granula
pri bojenju s hematoksilin eozinom. Kombinirano bojenje alciansko plavilo - periodično bojenje kiselinom
po Schiffu nije pokazalo značajnu varijaciju s obzirom na korišteni fiksativ i ukazuje da granule mastocita
štuke sadrže kisele, ali ne i neutralne mucine.

Abstract (english)

Piscine mast cells are also known as eosinophilic granular cells/eosinophilic granule cells.
Metachromasia of their granules is not often evident. According to the available literature, the cause may
be the composition of granules and/or their possible solubility after the use of aqueous fixatives. Staining
properties of mast cell granules show great differences, depending upon the fish species, the stage of the
life cycle and the health status.
The aim of this study was to test the suitability of certain fixatives in combination with staining
methods for visualization of mast cells in the northern pike (Esox lucius) collected from the Mrežnica River.
Paraffin blocks with different organs (digestive tract, pancreas and liver) from three northern pike were
used for this study. The sampled organs were fixed in four different fixatives (10 % neutral formalin,
Bouin's fixative, 100 % ethanol, and Carnoy's fixative) and stained with various histological and
histochemical methods (hematoxylin eosin, toluidine blue, Alcian blue - periodic Schiff acid staining, Alcian
blue - safranin, Verhoeff-Van Giesen).
With all of the fixatives and toluidine blue used, it was not possible to obtain metachromasia of
cytoplasmic granules. By using 10 % neutral formalin and toluidine blue, it is possible to obtain a clear
visualization of mast cells with orthochromatically stained granules. 100 % ethanol is not suitable fixative
for the intestine, while Carnoy's and Bouin's fixatives proved to be suitable for obtaining pronounced
eosinophilia of granules when stained with hematoxylin eosin. Alcian blue - periodic acid Schiff staining
showed no variation with respect to the fixative used. This indicates that the mast cell granules of northern
pike contain acidic but not neutral mucins.

• Keywords: mastociti; štuka; EGC; histološke i histokemijske metode
• Keywords (english): mast cells; pike; EGC; histology; histochemistry
• Language: croatian
• URN:NBN: urn:nbn:hr:178:572164
• Study programme: Title: Veterinary Medicine; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: doktor/doktorica veterinarske medicine (doktor/doktorica veterinarske medicine)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-01-05 09:23:38