Sample preparation and data collection for Serial Block Face Scanning Electron Microscopy of Mammalian Cell Monolayers v2
Year: 2024
Type: preprint
Abstract: Serial block face scanning electron microscopy (SBF-SEM) is a volume EM technique used to study biological samples including tissue and cellular samples. Here we describe detailed protocols for two sample embedding methods for in vitro tissue culture cells intended to be studied using SBF-SEM. The first protocol focuses on cell pellet embedding and the second on en face embedding. En face embedding can be combined with light microscopy, and this CLEM workflow can be used to identify specific biological events in a light microscope, which can then be imaged using SBF-SEM. We systematically outline the steps necessary to fix, stain, embed and image adherent tissue culture cell monolayers by SBF-SEM. Users experienced with electron microscopy sample preparation methodology will be able to complete this protocol in 10-11 days from initial seeding of cells in tissue culture to image acquisition.
Institutions: NYU Langone Health, New York University
Language: English
Subfield: Structural Biology
Domain: Life Sciences
Open Access status: green
Award: Cancer Center Support Grant
OpenAlex ID: W4392787574