在Cumulus-Oocyte复合体中成像跨区域投影:挑战和解决方案†
Mathilde Marchais1,2,3, Alexandre Bastien4, Elolo Karen Nenonene1,2,3
1Département des sciences animales, Faculté des sciences de l'agriculture et de l'alimentation, Université Laval, Québec, Qc, Canada.
Biology of reproduction
|September 5, 2025
概括
由于光学偏差, 对卵细胞进行深度3D成像是具有挑战性的. 优化样本准备和对焦设置可显著改善像跨区域投射 (TZP) 这样的细结构的成像.
科学领域:
- 生殖生物学
- 显微镜和成像技术
- 细胞生物学
背景情况:
- 对卵细胞进行深度3D成像面临诸如深度依赖的影子和光学偏差等挑战.
- 器官和脂质滴的折射率不匹配阻碍了精细结构的清晰可视化,包括跨区域投射 (TZPs).
研究的目的:
- 对比各种样本准备和共聚焦显微镜设置以改善卵细胞成像.
- 确定克服积卵细胞复合体 (COC) 深度3D成像的光学限制的方法.
主要方法:
- 使用固定胚胎的清除协议来证明深度限制.
- 清除与未清除标本的图像比较.
- 研究了累积密度,固定协议和透对图像质量的影响.
- 优化了对焦参数和评估了安装介质和幻灯片组装.
主要成果:
- 清除,脂质液滴去除和介质协调改善了染色和分布.
- 轻柔的剥离和低固定降低了图像噪声.
- 提顿透增强了抗体的透,从而有效地标记了TZP的蛋白质.
- 最佳的对焦设置和幻灯片组装对于样品分辨率至关重要.
结论:
- 样本的准备和对焦设置对深度3D卵细胞成像的质量产生重大影响.
- 优化的协议增强了积卵细胞复合体内TZP等细结构的可视化.
- 这些发现为生殖生物学和显微镜研究人员提供了实用解决方案.
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