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Farklı Fiksasyon Protokolleri ile Sperm Kromatin Kondansasyon Anomalisinin Değerlendirilmesi

Evaluation of Sperm Decondansation Anomalies with Different Fixation Protocols

Journal Name:

  • Uludağ Üniversitesi Tıp Fakültesi Dergisi

Publication Year:

  • 2006

Key Words:

Keywords (Original Language):

Author NameUniversity of Author
Abstract (2. Language): 
The integrity of sperm chromatin may play an important role for fertilization success, particularly in ICSI applications. Decreased nuclear chromatin condensation is assessed by using stain and fluorochroms such as aniline blue, acridine orange and chromamycine A3 (CMA3). Aniline blue is a stain that evaluates sperm morphology together with chromatin condensation anomalies. Fluorochroms are suggested that they are the most reliable stain for assessment of correlation between condensation anomalies and fertilization success. There are used different fixation protocols for these staining methods. .In this study, it is aimed to find the most practical and brief fixation protocole for evaluation of chromatin condensation. A total of 5 semen samples prepared by swim-up technic were fixed with Carnoy’s solution for 5 minutes at 4ºC or 2 hours at room temperature or 3% glutaraldehite for 20 minutes at room temperature. Aniline blue, acridine orange and CMA3 were applied on slides fixed with different fixatives. No staining quality difference was observed between groups. Our results showed that fixation with Carnoy’s solution for 5 minutes at 4ºC is satisfactory for all of the staining methods.
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Abstract (Original Language): 
Sperm kromatininin bütünlüğü, özellikle ICSI uygulamalarında, fertilizasyon başarısında önemli rol oynar. Nüklear kromatin kondansasyonundaki azalış aniline blue, acridine orange, chromamycine A3 (CMA3) gibi boya ve fluorokromlarla değerlendirilir. Aniline blue, kondansasyon anomalisi ile birlikte morfolojisini de değerlendiren bir boyadır. Fluorokromlar fertilizasyon başarı hızı ile kondansasyon anomalisi arasındaki korelasyonu saptamada en güvenilir boyalar olarak bildirilmiştir. Kısıtlı miktarda olan ICSI materyaline uygulanan bu boyalarda farklı fiksasyon protokolleri uygulanır. Bu çalışmada kromatin kondansasyonunu değerlendirmede uygulanan boyama metotlarında kullanılan en kısa ve pratik fiksasyon protokolünü bulmak amaçlandı. Toplam 5 hastanın swim-up yöntemi ile hazırlanan semen örnekleri Carnoy’un solüsyonunda 4ºC’de 5 dk, oda sıcaklığında 2 saat ve % 3’lük glutaraldehitte 20 dk fikse edildi.Farklı protokollerle fikse edilen preparatlara aniline blue, acridine orange ve CMA3 boyamaları uygulandı. Boyanma kalitesi açısından gruplarda fark görülmedi. Her üç boyamada da Carnoy’un solüsyonunda 4ºC’de 5 dk fiksasyonun yeterli olduğu görüşüne varıldı.
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  • 2
55-59
  • Turkish

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