AUTHOR=Merchant-Larios Horacio , Giraldo-Gomez David M. , Castro-Dominguez Adriana , Marmolejo-Valencia Alejandro 

TITLE=Light and focused ion beam microscopy workflow for resin-embedded tissues

JOURNAL=Frontiers in Cell and Developmental Biology

VOLUME=11

YEAR=2023

URL=https://www.frontiersin.org/journals/cell-and-developmental-biology/articles/10.3389/fcell.2023.1076736

DOI=10.3389/fcell.2023.1076736

ISSN=2296-634X

ABSTRACT=<p>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.</p>