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

Öz

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.

Anahtar Kelimeler

• 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

Öz

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.

Anahtar Kelimeler

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

Kaynakça

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