![]() Volume scanning EMs (SEMs) have opened the possibility of visualizing large volumes ( Peddie and Collinson, 2014 Titze and Genoud, 2016). The blooming of new technologies over the last decade has dramatically increased the value of EM for cell biology. ![]() ![]() Using this method, we acquired volumes of specific single cells within large tissues such as 3D cultures of mouse mammary gland organoids, tracheal terminal cells in Drosophila melanogaster larvae, and ovarian follicular cells in adult Drosophila, discovering ultrastructural details that could not be appreciated before. Laser branding is used to create landmarks on the block surface to position the FIB-SEM acquisition. The strategy relies on fluorescence preservation during sample preparation and targeted trimming guided by confocal maps of the fluorescence signal in the resin block. Here, we provide a workflow to target FIB-SEM acquisition of fluorescently labeled cells or subcellular structures with micrometer precision. While they allow automated data acquisition, precise targeting of volumes of interest within a large sample remains challenging. Today, scanning EM (SEM) methods such as focused ion beam (FIB)–SEM are frequently used for vEM analyses. Therefore, volume EM (vEM) is often crucial for correct interpretation of ultrastructural data. ![]()
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