Periferik Sinirlerin Histomorfometrik Analizinde TT-MMS Programının Güvenilirliğinin Karşılaştırmalı Olarak İncelenmesi

Deneysel çalışmalarda periferik sinirin akson çapı, miyelinli sinir lifi çapı ve miyelinli sinir için iç çap/dış çap oranı (g-ratio), demiyelinizasyon, remiyelinizasyon ve sinir onarımının değerlendirilmesinde görüntü analiz programlarından yararlanılmaktadır. Bu nedenle, morfometrik değerlendirmeye sıklıkla gereksinim duyulan periferik sinir çalışmaları hedeflenerek, laboratuvarımızda yarı otomatik bir görüntü analiz programı geliştirilmiştir. Çalışmamızda, geliştirdiğimiz görüntü analiz yazılımının performansının, serbest erişimli çok amaçlı diğer bir yazılım ile güvenilirlik yönünden karşılaştırılması amaçlanmıştır. Farelerin genel anestezi altında siyatik sinirlerinden alınan doku örnekleri gluteraldehit solüsyonuna alınmıştır. Rutin elektron mikroskop takip yöntemlerinden geçirilerek epoksi resin (epoxy resin) gömülen dokulardan yarı ince kesitler alınmış ve toluidin mavisi ile boyanmıştır. Kesitler dijital kameralı ışık mikroskopta incelenmiş ve X100’lük objektifte fotoğraflanmıştır. Mikrograflar laboratuvarımızda geliştirilen Tantuna Morfometrik Ölçüm Sistemi (TanTuna Morphometric Measuring System; TT-MMS) yazılımı ile analiz edilmiştir. Seçilen iki adet ışık mikrograftan rastgele seçilen 200 adet miyelinli akson kesitinin ölçümleri program kullanılarak yapılmıştır. Aynı ölçümler ImageJ programı ile tekrarlanmış ve her iki programla elde edilen veriler istatistiksel olarak karşılaştırılmıştır. Ek bir donanıma ihtiyaç duymadan her iki yazılımın da güvenilir olduğu ve birbirleriyle uyumlu ölçümler yaptıkları saptanmıştır (p

Anahtar Kelimeler:

akson çapı, g-ratio, ImageJ, periferik sinir, TT-MMS, axon diameter

Comparative Analysis of the Reliability of TT-MMS Program in Peripheral Nerves Histomorphometric Measurements

Image analysing programs are used in peripheral nerve studies to measure axon diameter, myelinated nerve diameter and the ratio of inner axonal diameter to outer diameter (g-ratio), demyelination, remyelination and nerve repair. So, a semi-automatic, image analysis program was developed in our laboratory by targeting peripheral nerve studies which morphometric evaluation is frequently required. It is aimed to compare the performance and reliability of this program with another free access multipurpose software. Tissue samples from sciatic nerves of mice were taken into gluteraldehyde solution under general anesthesia. After routine electron microscopic processing method, semi-thin sections were taken from epoxy resin embedded blocks and stained with toluidine blue. Sections were examined and photographed under the light microscope at X100 magnification. Micrographs were analyzed using the Tantuna Morphometric Measuring System (TT-MMS) software developed in our laboratory. Measurements of the randomly selected 200 myelinated axon sections from two selected light micrograph were performed using the program. The measurements were repeated with ImageJ program and data obtained using both programs were compared statistically. We determined that both softwares are reliable and measurements compatible with each other (p

Keywords:

axon diameter, g-ratio, ImageJ, peripheral nerve, TT-MMS,

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