[对从刺激周期获得的人类卵母细胞减数分裂纺锤体的侵入性和非侵入性分析：初步结果]
[Invasive and noninvasive analysis of the meiotic spindle of human oocytes obtained from stimulate cycles: preliminary results].
作者信息
Dib Luciana Azôr, da Broi Michele Gomes, de Araújo Maria Cristina Picinato Medeiros, Giorgenon Roberta Cristina, Ferriani Rui Alberto, Navarro Paula Andrea de Albuquerque Sales
机构信息
Departamento de Ginecologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brasil.
出版信息
Rev Bras Ginecol Obstet. 2012 Nov;34(11):524-9. doi: 10.1590/s0100-72032012001100008.
PURPOSE
To evaluate the concordance between polarization microscopy and confocal microscopy techniques in the evaluation of the meiotic spindle of human oocytes matured in vivo.
METHODS
Prospective study that evaluated oocytes with the first polar extruded body obtained from infertile women who had undergone ovarian stimulation for intracytoplasmic sperm injection. The oocytes with the first polar extruded body were evaluated by polarization microscopy and were then immediately fixed and stained for microtubule and chromatin evaluation by high-performance confocal microscopy. We determined the correlation of polarization microscopy with confocal microscopy in the detection of meiotic oocyte anomalies, and we also evaluated the percentage of oocytes with a visible and non-visible cell spindle by polarization microscopy and with meiotic normality and abnormalities by confocal microscopy. Confidence intervals, Kappa's index and concordance between the methodologies were calculated, considering immunofluorescence microscopy analysis as the golden-standard for evaluating normal spindle and oocyte chromosome distribution.
RESULTS
We observed that 72.7% of metaphase II oocytes with a nonvisible meiotic spindle by polarization microscopy showed no meiotic abnormalities by confocal analysis and 55.6% of metaphase II oocytes with a visible meiotic spindle by polarization microscopy were found to be abnormal oocytes by the confocal analysis. Only 44.4% of oocytes with a visible meiotic spindle by polarization microscopy were found to be normal by confocal analysis. Concordance between the methods was 51.1% (Kappa: 0.11; 95%CI -0.0958 - 0.319).
CONCLUSIONS
The low correlation between polarization microscopy and confocal microscopy in the assessment of oocyte meiotic spindle suggests that visualization of the meiotic spindle of human oocytes at metaphase II by polarization microscopy is not a good indicator of oocyte meiotic normality.
目的
评估偏振显微镜和共聚焦显微镜技术在评估体内成熟的人类卵母细胞减数分裂纺锤体方面的一致性。
方法
前瞻性研究，评估从接受卵巢刺激以进行胞浆内单精子注射的不孕妇女中获得的带有第一极体的卵母细胞。对带有第一极体的卵母细胞进行偏振显微镜评估，然后立即固定并染色，通过高性能共聚焦显微镜进行微管和染色质评估。我们确定了偏振显微镜与共聚焦显微镜在检测卵母细胞减数分裂异常方面的相关性，还评估了通过偏振显微镜观察到可见和不可见细胞纺锤体的卵母细胞百分比，以及通过共聚焦显微镜观察到的减数分裂正常和异常的卵母细胞百分比。计算了置信区间、卡帕指数以及两种方法之间的一致性，将免疫荧光显微镜分析视为评估正常纺锤体和卵母细胞染色体分布的金标准。
结果
我们观察到，通过偏振显微镜观察到减数分裂纺锤体不可见的中期II卵母细胞中，72.7%经共聚焦分析显示无减数分裂异常；通过偏振显微镜观察到减数分裂纺锤体可见的中期II卵母细胞中，55.6%经共聚焦分析发现为异常卵母细胞。通过偏振显微镜观察到减数分裂纺锤体可见的卵母细胞中，只有44.4%经共聚焦分析发现为正常。两种方法之间的一致性为51.1%（卡帕：0.11；95%置信区间 -0.0958 - 0.319）。
结论
偏振显微镜和共聚焦显微镜在评估卵母细胞减数分裂纺锤体方面的低相关性表明，通过偏振显微镜观察中期II人类卵母细胞的减数分裂纺锤体并非卵母细胞减数分裂正常的良好指标。
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