Comparison of Flow Cytometric Analysis and Eosin-Nigrosin Staining Methods for Determining some Morphological Characteristics of Bull Epididymal Spermatozoa

AKYOL, NUMAN; AZKUR, Ahmet Kursat; EKİCİ, Hüsamettin

Comparison of Flow Cytometric Analysis and Eosin-Nigrosin Staining Methods for Determining some Morphological Characteristics of Bull Epididymal Spermatozoa

Numan AKYOL, (Kırıkkale Üniversitesi, Veteriner Fakültesi, Dölerme ve Suni Tohumlama Anabilim Dalı, Kırıkkale-Türkiye)

Ahmet Kürşat AZKUR, (Kırıkkale Üniversitesi, Veteriner Fakültesi, Viroloji Anabilim Dalı, Kırıkkale, Türkiye)

Hüsamettin EKİCİ (Kırıkkale Üniversitesi, Veteriner Fakültesi, Farmakoloji ve Toksikoloji Anabilim Dalı, Kırıkkale Türkiye)

Kafkas Üniversitesi Veteriner Fakültesi Dergisi

2 0

Yıl: 2015 Cilt: 21 Sayı: 5 Sayfa Aralığı: 659 - 664 Metin Dili: Türkçe İndeks Tarihi: 29-07-2022

Comparison of Flow Cytometric Analysis and Eosin-Nigrosin Staining Methods for Determining some Morphological Characteristics of Bull Epididymal Spermatozoa

Öz:

Bu çalışmanın amacı, epididimal kaynaklı boğa spermatozoonlarının taze ve dondurulup çözdürüldükten sonra akım sitometri (FlowCytometri) yöntemiyle analiz edilmesi, nekroz/apoptoz düzeylerinin incelenmesi ve klasik boyama yöntemi olan eozin-nigrozinboyama metoduyla kıyaslanması amaçlanmıştır. Bu amaçla yerel mezbahada kesilen 14 boğanın epididimislerinden elde edilenspermatozoonlar kullanılmıştır. Canlı spermatozoa, toplam apoptotik, nekrotik ve erken nekrotik spermatozoa düzeyleri akım sitometriyöntemiyle tespit edilmiştir ve bu amaçla AnnexinV+PI (Propidium İyodür) floresan boyama yöntemi kullanılmıştır. Taze ve çözdürülmüşsperma içerisindeki, ölü spermatozoa ve protoplazmik damlacık taşıyan spermatozoonların oranları eozin-nigrozin boyama yöntemiile belirlenmiştir. Taze spermada, akım sitometri ile ölü spermatozoon sayısı eozin-nigrozin boyamada bulunana göre daha az olduğugörülmüştür (P<0.05). Protoplazmik damlacık gibi bazı morfoloj ik parametrelerin mikroskopik muayenelerle belirlenebildiği ancakölüm sürecine giren spermatozoonların alt kategorilerinin (apoptotik, nekrotik, erken nekrotik) akım sitometri ile net olarak tespitedilebildiği görülmüştür. Sonuç olarak, klasik yöntemlere göre spermatozoa canlılığı tespitinde akım sitometri yönteminin klasikboyama yöntemlerine destek olarak ve daha etkin bir şekilde kullanılabileceği düşünülmüştür.

Anahtar Kelime:

Konular: Veterinerlik

Epididimal boğa Spermatozoonlarının Bazı Morfolojik Özelliklerinin Belirlenmesi Amacıyla Akım Sitometri ve Eozin-Nigrozin Boyama Yöntemlerinin Karşılaştırılması

Öz:

The aim of this study is to investigate necrosis and apoptosis in epididymal bull spermatozoa before freezing and after thawing usingthe flow cytometric method and to compare this with eosin-nigrosin dyeing, which is the conventional method used in assessing ofspermatozoa. The testicles from fourteen bulls at local slaughterhouse were used for this study. The proportions of live spermatozoa,total apoptotic, necrotic and early necrotic spermatozoa levels were observed via flow cytometry. Annexin V/PI fluoruscence dyeingwas used to investigate the proprotions of apoptotic, necrotic, early necrotic and live spermatozoa for flow cytometry. The proportionof dead spermatozoa and protoplasmic droplets were determined using the eosin-nigrosin conventional dyeing method in fresh andfrozen-thawed spermatozoa. The average dead spermatozoa count with flow cytometry was less than with the eosin-nigrosin method(P<0.05). Some morphological characteristics such as protoplasmic droplets could be determined with the eosin-nigrosin method;however, sperm subpopulations entering the death process (apoptotic, necrotic and early necrotic) could be defined clearly onlywith the flow cytometric method. As a result, combination of eosin-nigrosin dyeing method and flow cytometric analysis of spermmorphological evaluation could give better results of bull epididymal semen in comparison to eosin-nigrosin dyeing method alone.

Anahtar Kelime:

Konular: Veterinerlik

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

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APA	AKYOL N, AZKUR A, EKİCİ H (2015). Comparison of Flow Cytometric Analysis and Eosin-Nigrosin Staining Methods for Determining some Morphological Characteristics of Bull Epididymal Spermatozoa. , 659 - 664.
Chicago	AKYOL NUMAN,AZKUR Ahmet Kursat,EKİCİ Hüsamettin Comparison of Flow Cytometric Analysis and Eosin-Nigrosin Staining Methods for Determining some Morphological Characteristics of Bull Epididymal Spermatozoa. (2015): 659 - 664.
MLA	AKYOL NUMAN,AZKUR Ahmet Kursat,EKİCİ Hüsamettin Comparison of Flow Cytometric Analysis and Eosin-Nigrosin Staining Methods for Determining some Morphological Characteristics of Bull Epididymal Spermatozoa. , 2015, ss.659 - 664.
AMA	AKYOL N,AZKUR A,EKİCİ H Comparison of Flow Cytometric Analysis and Eosin-Nigrosin Staining Methods for Determining some Morphological Characteristics of Bull Epididymal Spermatozoa. . 2015; 659 - 664.
Vancouver	AKYOL N,AZKUR A,EKİCİ H Comparison of Flow Cytometric Analysis and Eosin-Nigrosin Staining Methods for Determining some Morphological Characteristics of Bull Epididymal Spermatozoa. . 2015; 659 - 664.
IEEE	AKYOL N,AZKUR A,EKİCİ H "Comparison of Flow Cytometric Analysis and Eosin-Nigrosin Staining Methods for Determining some Morphological Characteristics of Bull Epididymal Spermatozoa." , ss.659 - 664, 2015.
ISNAD	AKYOL, NUMAN vd. "Comparison of Flow Cytometric Analysis and Eosin-Nigrosin Staining Methods for Determining some Morphological Characteristics of Bull Epididymal Spermatozoa". (2015), 659-664.

APA	AKYOL N, AZKUR A, EKİCİ H (2015). Comparison of Flow Cytometric Analysis and Eosin-Nigrosin Staining Methods for Determining some Morphological Characteristics of Bull Epididymal Spermatozoa. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, 21(5), 659 - 664.
Chicago	AKYOL NUMAN,AZKUR Ahmet Kursat,EKİCİ Hüsamettin Comparison of Flow Cytometric Analysis and Eosin-Nigrosin Staining Methods for Determining some Morphological Characteristics of Bull Epididymal Spermatozoa. Kafkas Üniversitesi Veteriner Fakültesi Dergisi 21, no.5 (2015): 659 - 664.
MLA	AKYOL NUMAN,AZKUR Ahmet Kursat,EKİCİ Hüsamettin Comparison of Flow Cytometric Analysis and Eosin-Nigrosin Staining Methods for Determining some Morphological Characteristics of Bull Epididymal Spermatozoa. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, vol.21, no.5, 2015, ss.659 - 664.
AMA	AKYOL N,AZKUR A,EKİCİ H Comparison of Flow Cytometric Analysis and Eosin-Nigrosin Staining Methods for Determining some Morphological Characteristics of Bull Epididymal Spermatozoa. Kafkas Üniversitesi Veteriner Fakültesi Dergisi. 2015; 21(5): 659 - 664.
Vancouver	AKYOL N,AZKUR A,EKİCİ H Comparison of Flow Cytometric Analysis and Eosin-Nigrosin Staining Methods for Determining some Morphological Characteristics of Bull Epididymal Spermatozoa. Kafkas Üniversitesi Veteriner Fakültesi Dergisi. 2015; 21(5): 659 - 664.
IEEE	AKYOL N,AZKUR A,EKİCİ H "Comparison of Flow Cytometric Analysis and Eosin-Nigrosin Staining Methods for Determining some Morphological Characteristics of Bull Epididymal Spermatozoa." Kafkas Üniversitesi Veteriner Fakültesi Dergisi, 21, ss.659 - 664, 2015.
ISNAD	AKYOL, NUMAN vd. "Comparison of Flow Cytometric Analysis and Eosin-Nigrosin Staining Methods for Determining some Morphological Characteristics of Bull Epididymal Spermatozoa". Kafkas Üniversitesi Veteriner Fakültesi Dergisi 21/5 (2015), 659-664.