Comparative Examination of Commonly Used Some Fixatives with Routine Histochemical Staining’s for The Optimal Histological Appearance in The Gill Tissue of Zebrafish

Year 2019, Volume: 12 Issue: 2, 158 - 167, 30.06.2019

Aykut Ulucan , Hayati Yüksel , Muhammed Bahaeddin Dörtbudak Seda Yakut

https://doi.org/10.30607/kvj.526779

Cited By: 2

Abstract

Histopathological studies related to Zebrafish which
are used as a model organism in researching the pathogenesis of many diseases
increase day by day. Because of the increasing importance of zebrafish
research, experiments on this animal model are more prominent and gill tissue
is frequently examined in various disease models using zebrafish. As it is
known, at the end of the experimental animal model studies, if the
histopathological examination is to be done, the tissues should be fixed. The
purpose of fixation is to keep cellular and extracellular components in vivo as
much as possible. Therefore, the choice of fixation methods and fixatives has a
significant effect on tissue processing. In this study, we aimed to optimize
fixation techniques and staining protocols for producing ideal slides and
histological images of gill tissue in zebrafish. In our study, to determine the
optimal histology in the zebrafish-gill tissue, the tissues were fixed with
Bouin’s, Carnoy’s, 10% neutral buffered formalin (NBF), Davidson’s, and
Dietrich’s solutions. Following the routine tissue processing, the sections
were stained with Hematoxylin and Eosin (H&E) and Masson’s Trichrome (MT)
stains. Consequently, tissue morphology was best preserved with Bouin’s and NBF
solutions. The best results in H&E stain were obtained from tissues fixed
with NBF solution, while Dietrich's solution fixation was found to be ideal for
MT staining.

Keywords

Fixatives, Hematoxylin and Eosin, Masson’s Trichrome, Zebrafish, Gill

Zebra Balığı Solungaç Dokusunda Optimal Histolojik Görünüm için Rutin Histokimyasal Boyama ile Sık Kullanılan Fiksatiflerin Karşılaştırmalı Olarak İncelenmesi

Abstract

Birçok hastalığın patogenezinin araştırılmasında model
bir organizma olarak kullanılan Zebra balığı ile ilgili histopatolojik
çalışmalar gün geçtikçe artmaktadır. Zebra balığı araştırmalarının artan önemi
nedeniyle, bu hayvan modelindeki deneyler daha önemlidir ve solungaç dokusu
zebra balığı kullanılan çeşitli hastalık modellerinde sıklıkla incelenir.
Bilindiği üzere, deneysel hayvan modeli çalışmalarının sonunda, eğer
histopatolojik inceleme yapılacaksa, dokular fikse edilmelidir. Fiksasyonun
amacı, hücresel ve hücre dışı bileşenleri mümkün olduğunca in vivo halde
korumaktır. Bu nedenle, fiksasyon yöntemlerinin ve fiksatiflerin seçiminin doku
takibi üzerinde önemli bir etkisi vardır. Bu çalışmada, zebra balığı solungaç dokusundan
ideal preparatlar ve  histolojik
görüntüler elde etmek için fiksasyon tekniklerini ve boyama protokollerini
optimize etmek amaçlanmıştır. Çalışmamızda, zebra balığı-solungaç dokusundaki
ideal histolojiyi belirlemek için, dokular, Bouin, Carnoy, %10’luk nötral
tamponlu formaldehit, Davidson ve Dietrich solüsyonları ile fikse edilmiştir.
Rutin doku işleminin ardından, kesitler Hematoksilen ve Eosin (H&E) ve
Masson Trikrom (MT) boyaları ile boyandı. Sonuç olarak, doku morfolojisi en iyi
Bouin ve NBF solüsyonları ile korunmuştur. H&E boyamasında en iyi sonuçlar
NBF çözeltisi ile fikse edilmiş dokulardan elde edilirken, Dietrich solüsyonu
fiksasyonunun MT boyaması için ideal olduğu bulundu.

Keywords

Fiksatifler, Hematoxylin ve Eosin, Masson Trikrom, Zebra balığı, Solungaç

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