Farklı boyamaların çeşitli dokuların parafin ve yarı ince epon kesitlerine uygulanmasının ışık mikroskobik düzeyde karşılaştırılması
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Date
2021-02-15
Authors
Akbaş, Ayşe
Journal Title
Journal ISSN
Volume Title
Publisher
Bursa Uludağ Üniversitesi
Abstract
Histolojide mikroskobik değerlendirme amacıyla alınan doku örnekleri, parafine veya plastik ortamlara gömülür. Plastik yarı ince kesitler, ışık mikroskobu ile elektron mikroskobu arasında bir köprü görevi görür. Bu kesitler parafinden daha ince, ince kesitlerden daha kalındır. Bu sayede ışık mikroskopide yarı ince kesitler, parafine göre daha ileri detay sağlar. Ancak plastik kesitlerin çoğu hidrofobiktir ve birçok boya ile boyanmazlar. Bu çalışmada; parafin ve epon kesitlerde daha çok doku komponentinin ayrımına imkan verecek boyama metotlarının geliştirilmesi amaçlanmıştır. Çalışmada; çeşitli doku örnekleri ikili fiksasyon sonrası Epon ile bloklandı. Aynı dokuların diğer grubu, rutin takip sonrası parafin ile bloklandı. Toluidine blue, hematoksilen, eozin, asit fuksin, metil green ve nötral red boyaları hem tekli, hem ikili kombinasyonlar şeklinde her iki kesit grubunda denendi. Ayrıca elastik, retiküler ve kollajen lifler ile mitokondri boyamaları da çalışıldı. Epon kesitlere boyaların penetrasyonunu kolaylaştırmak amacıyla, üç ayrı etching ajanı (sodyum hidroksit, periyodik asit ve aseton) uygulandı. Sonuçlar ışık mikroskobunda değerlendirildi. Eponun uzaklaştırılmasında aseton en başarılı bulundu. Epon kesitlerde tekli boyamalarda en iyi sonuçlar toluidine blue ile sağlandı. Ayrıca asit fuksin ve metil green de olumlu bulundu. Tekli boyamalar parafin kesitlerde başarılı bulunmadı. Epon kesitlerde ikili boyamalarda; toluidine blue-eozin, toluidine blue-asit fuksin, toluidine blue-metil green ve asit fuksin-toluidine blue kombinasyonlarında başarılı sonuçlar elde edildi. Parafin kesitlerde hematoksilen-eozin ile en iyi sonuçlar alındı. Ayrıca; toluidine blue-eozin ve asit fuksin-metil green kombinasyonlarından da yararlanılabileceği düşünüldü. Parafin ve epon kesitlerde dört özel boyama ile elastik, retiküler ve kollajen lifler ile mitokondri demonstrasyonunda da başarılı sonuçlar elde edildi. Olumlu bulunan boyamaların rutinde kullanılabileceği sonucuna varıldı.
Tissue samples received for microscopic evaluation in histology are embedded in paraffin or plastic media. Plastic semithin sections serve as a connection between light and electron microscopies. These sections are thinner than paraffin sections, and thicker than thin sections. This helps semi-thin sections in light microscopy provide more advanced details than paraffin sections do. Most plastic sections are hydrophobic, however, and cannot be stained with many stains. In this study, the aim is to develop staining methods that allow more tissue components to be differantiated in paraffin and epon sections. Various tissue samples were subjected to double fixation and epon blocking in the study. Another group of the same tissues was subjected to routine tissue processing and paraffin blocking. Toluidine blue, hematoxylin, eosin, acid fuchsin, methyl green, and neutral red stains were tested in both section groups as single stains or dual combinations. Elastic, reticular, and collagen fiber stains and mitochondrial stains were tested additionally. Three different etching agents (sodium hydroxide, periodic acid, and acetone) were applied to facilitate the penetration of stains into the epon sections. The results were evaluated under a light microscope. Acetone was found to be the most successful in removing epon. The best results in epon sections with single stains were attained with toluidine blue. Acid fuchsin and methyl green were also found satisfactory. Single stains were not successful in paraffin sections. Among the dual stainings in epon sections, toluidine blue-eosin, toluidine blue-acid fuchsin, toluidine blue-methyl green, and acid fuchsin-toluidine blue combinations yielded successful results. The best results in paraffin sections were obtained with hematoxylin-eosin. Toluidine blue-eosin and acid fuchsin-methyl green combinations were also considered useful. Four special stainings yielded good results in demonstration of elastic, reticular, and collagen fibers and mitochondria in paraffin and epon sections. It was concluded that the stainings that are found successful can be used routinely.
Tissue samples received for microscopic evaluation in histology are embedded in paraffin or plastic media. Plastic semithin sections serve as a connection between light and electron microscopies. These sections are thinner than paraffin sections, and thicker than thin sections. This helps semi-thin sections in light microscopy provide more advanced details than paraffin sections do. Most plastic sections are hydrophobic, however, and cannot be stained with many stains. In this study, the aim is to develop staining methods that allow more tissue components to be differantiated in paraffin and epon sections. Various tissue samples were subjected to double fixation and epon blocking in the study. Another group of the same tissues was subjected to routine tissue processing and paraffin blocking. Toluidine blue, hematoxylin, eosin, acid fuchsin, methyl green, and neutral red stains were tested in both section groups as single stains or dual combinations. Elastic, reticular, and collagen fiber stains and mitochondrial stains were tested additionally. Three different etching agents (sodium hydroxide, periodic acid, and acetone) were applied to facilitate the penetration of stains into the epon sections. The results were evaluated under a light microscope. Acetone was found to be the most successful in removing epon. The best results in epon sections with single stains were attained with toluidine blue. Acid fuchsin and methyl green were also found satisfactory. Single stains were not successful in paraffin sections. Among the dual stainings in epon sections, toluidine blue-eosin, toluidine blue-acid fuchsin, toluidine blue-methyl green, and acid fuchsin-toluidine blue combinations yielded successful results. The best results in paraffin sections were obtained with hematoxylin-eosin. Toluidine blue-eosin and acid fuchsin-methyl green combinations were also considered useful. Four special stainings yielded good results in demonstration of elastic, reticular, and collagen fibers and mitochondria in paraffin and epon sections. It was concluded that the stainings that are found successful can be used routinely.
Description
Keywords
Yarı ince epon kesit, Etching, Boyama, Işık mikroskopi, Semithin epon section, Staining, Light microscopy
Citation
Akbaş, A. (2021). Farklı boyamaların çeşitli dokuların parafin ve yarı ince epon kesitlerine uygulanmasının ışık mikroskobik düzeyde karşılaştırılması. Yayınlanmamış yüksek lisans tezi. Bursa Uludağ Üniversitesi Sağlık Bilimleri Enstitüsü.