Evaluation of nuclear condensation and spermatozoa morphology at the level of light microscope in different semen parameters

Year 2009, Volume: 40 Issue: 3, 111 - 117, 01.04.2009

Emine Aksoy Tahsin Murad Aktan Selçuk Duman Duygu Dursunoğlu Gökhan Cüce

Abstract

Aim: It was aimed to evaluate sperm DNA decondansation of oligoasthenoospermic and teratozoospermic patients at the level of light microscope. Material and Metods: Seminal plazma of oligoasthenoospermic (n: 20) and teratozoospermic (n: 25) samples were washed twice with PBS and then smear was prepared, fixation was done with metil alcohol. Staining was done with; heamatoxylin-eosin, toluidine blue, Giemsa and Wright stains. Under light microscopy 200 sperm was counted, condansated and decondansated ones were counted. Findings: Between four stains condensation/decondensation selection was done more easier with haematoxylin-eosin and toluidine stains. Statistical analyses did not show any difference on condensation evaluation between stains. There was no any significant difference for condensation ratio values between groups. Also comparasion between oligoasthenoospermic and teratozoospermic samples done with stains showed no any difference. But when safety area of graphic was considered (especially when teratozoospermic was compared to oligoasthenoospermic) toluidine stain showed a homogen appearance. Condensated sperm ratio was higher in teratozoospermic samples. Results: As a result toluidine blue is the most trustable stainjbr condensation evaluation. There was no any difference between four stains between oligoasthenoospermic and teratozoospermic samples. For intracytoplasmic sperm injection teratozoospermic samples are more reliable than oligoasthenoospermic samples.

Keywords

Microscopy, Polarization, Sperm Count, Oligospermia, Spermatozoa, Morphology

Farklı semen paramerrelerinde ışık mikroskobu düzeyinde spermatozoa morfolojisi ve nükleer kondansasyon değerlendirmesi

Abstract

Amaç: Oligozoospermili ve teratozoospermili hastalarda Işık Mikroskobu düzeyinde sperm morfolojisi ve sperm DNA kondansasyonunun değerlendirilmesi amaçlanmıştır. Gereç ve Yöntem: oligozoospermili (n: 20) ve teratozoospermili (n: 25) hastalardan alınan seminal plazma örnekleri PBS ile 2 kez yıkandı ve smear hazırlandı, metil alkol ile fikzasyonu hazırlandı. Hematoksilen-Eosin, Toluidin-Blue, Giemsa ve Wright özel histolojik boyaları ile boyandı. Işık mikroskobunda 200 sperm hücresi sayılıp kondanse ve dekondanse başa sahip olan spermatozoalar belirlendi. Bulgular: Bu dört boya ile yapılan yaymalarda kondanse spermler, Hematoksüen-Eozin ve Toluidin Blue boyası ile boyanmış preperatlarda daha kolay ve net görüldü. İstatistiksel olarak dört boya ile kondansasyon değerlendirmesinde hasta grupları arasında fark olmadığı görüldü. Ayrıca oligozoospermili ve teratozoospermili hasta grupları için boyalar kendi içinde karşılaştırıldığında fark olmadığı görüldü. Ancak güven aralığı grafiğine bakıldığında; teratozoospermili hastaların oligozoospermili hastalara göre özellikle de Toluidin blue ile boyanmış grubun belirgin ölçüde homojen dağılım göstermesi; teratozoospermili hasta grubunun daha güvenilir kondanse sperm sağladığını gösterdi Sonuç: Sperm kondansasyon değerlendirmesi için toluidin blue boyası en güvenilir boyalardan bindir. Oligozoospermili ve teratozoospermili hastalar arasında ve bu dört boya arasında sperm nükleer kondansasyonunda fark yoktur. Ancak intrastoplazmik sperm injeksiyonu için kondanse sperm seçimi yapılırken oligozoospermili hastalara göre teratozoospermili hastalar daha güvenilir sonuç verebilir.

Keywords

Mikroskopi, polarizasyon, Sperm sayısı, Oligospermi, Sperm, Morfoloji

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