Merchant-Larios Horacio, Giraldo-Gomez David M, Castro-Dominguez Adriana, Marmolejo-Valencia Alejandro
Instituto de Investigaciones Biomédicas Departamento de Biología Celular y Fisiología, Universidad Nacional Autónoma de México (UNAM), México City, México.
Carl Zeiss de México S. A. de C. V., Research Microscopy Solutions, , México City, Mexico.
Front Cell Dev Biol. 2023 Jan 25;11:1076736. doi: 10.3389/fcell.2023.1076736. eCollection 2023.
Although the automated image acquisition with the focused ion beam scanning electron microscope (FIB-SEM) provides volume reconstructions, volume analysis of large samples remains challenging. Here, we present a workflow that combines a modified sample protocol of the classical transmission electron microscope with FIB-SEM volume imaging. The proposed workflow enables efficient 3D structural surveys of rabbit ovaries collected at consecutive developmental stages. The precise trimming of the region of interest adds the time dimension to the volume, constructing a virtual 4D electron microscopy. We found filopodia-like processes emitted by oocyte cysts allowing contact between oocytes not previously observed.
尽管聚焦离子束扫描电子显微镜（FIB-SEM）的自动图像采集可提供体积重建，但对大样本进行体积分析仍具有挑战性。在此，我们提出了一种工作流程，该流程将经典透射电子显微镜的改良样本方案与FIB-SEM体积成像相结合。所提出的工作流程能够对在连续发育阶段采集的兔卵巢进行高效的三维结构研究。对感兴趣区域的精确修剪为体积增加了时间维度，构建了虚拟的四维电子显微镜。我们发现卵母细胞囊肿发出丝状伪足样突起，使得以前未观察到的卵母细胞之间能够接触。
