DURMUŞ BOLAT, SADULLAH BAHAR, EMRAH SUR, Muhammet L SELCUK, Sadettin TIPIRDAMAZ

Selective gray and white matter staining of the horse spinal cord

Medulla spinalis’te yer alan gri ve ak madde oranları diffuz ve kompresif omurilik hastalıklarının teşhisinde önemli klinik para- metrelerdendir. Bu parametrelerin tespitinde histolojik metotlar kullanılmasına rağmen, histolojik çalışmalarda doku büyüklüğüne bağlı bazı güçlüklüklerle karşılaşılmaktadır. Bu çalışmanın amacı, bahsedilen zorlukları aşmaya yönelik olası modifikasyonları değerlendirmektir. Çalışmada, Shetland pony ırkına ait C6 segmentinden sistematik rastgele örnekleme kuralına sadık kalınarak elde edilen 9 doku kesiti kullanıldı. Medulla spinalis’in dehidrasyon işlemi sırasında vakum uygulaması yapıldı. Dokuların parafin blokları hazırlandı, dokular 10 µm kalınlığında kesilerek farklı histolojik boyalar ile boyandı. Modifiye May - Grunwald - Giemsa (MMGG)’nın yer aldığı 6 farklı boyama metodunun boyama performansları karşılaştırıldı. Hematoxylin & eosin (H&E), May-Grunwald-Giemsa (MGG), Masson’s trichrome (MT), AgNORs, Kluver Barrera (KB) ve MMGG ile boyanan kesitler tek kör grup tarafından makroskopik ve mikroskopik olarak değerlendirildi. Boyaların performansları Likert skalası kullanılarak en kötü (1) en iyi (5) olmak üzere değerlendirildi. Vakum uygulamasının yetersiz doku dehidrasyonundan kaynaklanan problemleri ortadan kaldırdığı görüldü. MMGG, at medulla spinalis’inde gri ve ak madde ayrımında en başarılı boya olarak tespit edildi.

Anahtar Kelimeler:

omurilik, doku bilimi, eozin, giemsa boyaması, boyama, at

At omuriliğinin gri ve ak maddesinin seçici boyanması

The ratio of gray and white matter is an important clinical parameter in the diagnosis of diffuse and compressive diseases of the spinal cord. Although histological methods are used to determine this parameter, there are some difficulties encountered in histological studies related to tissue size. The aim of this study was to evaluate possible modifications to overcome these difficulties. In the study, nine tissue samples taken from the C6 segment of a female Shetland pony and selected by systematic random sampling were used. The dehydration process of the spinal cord of the horse was supported by applying a vacuum. Paraffin blocks were prepared and cut into 10 µm sections to be stained separately with the different staining methods. Six different staining methods, including Modified May - Grunwald - Giemsa (MMGG), were compared and used to image entire slides. The stains, Hematoxylin & eosin (H&E), May-Grunwald-Giemsa (MGG), Masson’s trichrome (MT), AgNORs, Kluver Barrera (KB) and MMGG, were evaluated macroscopically and microscopically by participants who were unaware of which staining methods had been used. The staining methods were scored from worst (1) to best (5) using a Likert scale. Vacuum application was found to reduce the difficulties related to inadequate tissue dehydration. MMGG was selected as the best staining method in differentiating gray and white matter in the spinal cord of the horse.

Keywords:

spinal cord, histology, eosine, Giemsa staining, staining, horses,

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