2015

Kedi ve Köpek Dokularının Farklı Fiksatiflerle Tespiti ve Histolojik Görünümlerinin Karşılaştırılması*

Tespit işlemi, hücre ve doku komponentlerinin ölüm sonrasında morfolojik özelliklerinin canlıdaki durumuna en yakın şekilde sabitleyip, histolojik olarak incelenmesine olanak tanır. Tespit işleminde kullanılan kimyasal ajanlara fik-satif denir. Yirminci yüzyıldan günümüze kadar rutin doku tespit işleminde formaldehit çözeltisi kullanılmaktadır. Her fiksatif doku tespit işlemi süresince doku histomorfolojisinin incelenmesi adına avantaj ve dezavantajlara sahiptir. Bu nedenle tespit işlemi için amaca yönelik doğru fiksatif ile çalışılması gerekir. Bu çalışmada hedef, 3 farklı fiksatif ile hazırlanan 4 çözeltinin, diğer faktörler optimum seviyede tutularak, doku tespit işlemi sürecinde nasıl etkilere sahip olduklarını belirlemek ve tespit özelliklerine göre en ideal kullanım amaçlarını ortaya çıkarmaktı. Bu amaçla, çalışmada doku tespit işlemi sırasında fiksatiflerin dezavantajlarından etkilenen ve hücre morfolojisinin incelenmesinde zorluklarla karşılaşılan dalak ve böbrek dokuları üzerinde çalışıldı. Yaklaşık 0.5 cm kalınlığındaki doku örnekleri hazırlanan %3.7`lik, %10`luk formaldehit ve Hollande çözeltilerinde 16 saat, Carnoy çözeltisinde ise 4 saat süre ile tespit edildi. Ru-tin doku takip işlemlerinden geçirildikten sonra alınan doku kesitleri H&E ile boyandı. Bu dokuların histomorfolojik ince-lemeleri yapıldığında hücre sitoplazma boyanması için en uygun fiksatifin %3.7`lik formaldehit çözeltisi olduğu, hücre çekirdek ve çekirdekçiklerinin morfolojisini ortaya koymada ise Carnoy ve Hollande çözeltilerin kullanımlarının ideal olduğu, ancak Carnoy çözeltisinin eritrositlerde ileri düzeyde lizise yol açtığı gözlendi. Sonuç olarak, doku tespit işle-minde %3.7`lik formaldehit çözeltisinin kullanımı ideal bulunurken, Carnoy ve Hollande çözeltilerinin ise hücre çekirdek-lerindeki histomorfolojik değişikliklerin detaylı incelenmesini gerektiren hastalıklarda etkili bir kullanım oluşturacakları ön görüldü.

Anahtar Kelimeler:

Carnoy, formaldehit, Hollande

Fixation and Comparative Histologic Evaluation of Cat and Dog Tissues using Different Fixatives

Fixation is a procedure that allows histological evaluation of cells and tissue components by fixing them to their closest morphological features in life-like state after death. Formaldehyde solution has been in use since the 20th century. During the tissue fixation, each fixatives represent their own advantages and disadvantages on the examina-tion of histomorphological properties of the tissues. So that, the fixation has to be performed with the appropriate fixa-tive depending on the research aim. The aim of this study was to determine the effects of four solutions, prepared by 3 different fixatives, on the tissue fixation process by keeping the other factors at an optimum level and to determine the most ideal use according to the fixation properties. For this purpose, the spleen and kidney tissues were studied, since they are generally affected by the disadvantages of fixatives during the tissue fixation process and lead to difficulties in examining cell morphology. Tissue samples of 0.5 cm thickness were fixed in the 3.7%, 10% formaldehyde and Hol-lande solutions for 16 hours and Carnoy solution for 4 hours. Thereafter, tissue samples were routinely proceeded, and sections were stained by H&E. The histomorphological examinations of these tissues revealed that the most suitable fixative for cytoplasmic staining was 3.7% formaldehyde solution, while the use of Carnoy and Hollande solutions was ideal in revealing the morphology of the nucleus and nucleolus. However the Carnoy solution caused high lysis in erythrocytes. As a result, it was concluded that the 3.7% formaldehyde solution could be ideally used in tissue fixation process, whereas Carnoy and Hollande solutions were seen to be effective in diseases that require detailed examina-tion of histomorphological changes in nucleus and nucleolus.

Keywords:

Carnoy fixation, formaldehyde, Hollande,

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