Precise targeting for 3D cryo-correlative light and electron microscopy volume imaging of tissues using a FinderTOP de Beer, Marit Daviran, Deniz Roverts, Rona Rutten, Luco Macías Sánchez, Elena R. Metz, Juriaan Sommerdijk, Nico Akiva, Anat Cryo-correlative light and electron microscopy (cryoCLEM) is a powerful strategy to high resolution imaging in the unperturbed hydrated state. In this approach fluorescence microscopy aids localizing the area of interest, and cryogenic focused ion beam/scanning electron microscopy (cryoFIB/SEM) allows preparation of thin cryo-lamellae for cryoET. However, the current method cannot be accurately applied on bulky (3D) samples such as tissues and organoids. 3D cryo-correlative imaging of large volumes is needed to close the resolution gap between cryo-light microscopy and cryoET, placing sub-nanometer observations in a larger biological context. Currently technological hurdles render 3D cryoCLEM an unexplored approach. Here we demonstrate a cryoCLEM workflow for tissues, correlating cryo-Airyscan confocal microscopy with 3D cryoFIB/SEM volume imaging. Accurate correlation is achieved by imprinting a FinderTOP pattern in the sample surface during high pressure freezing, and allows precise targeting for cryoFIB/SEM volume imaging. 2024-10-02T10:29:32Z 2024-10-02T10:29:32Z 2023-05-11 journal article de Beer, M. et. al. Commun Biol 6, 510 (2023). [https://doi.org/10.1038/s42003-023-04887-y] https://hdl.handle.net/10481/95421 10.1038/s42003-023-04887-y eng info:eu-repo/grantAgreement/EC/H2020/788982 info:eu-repo/grantAgreement/EC/H2020/101031624 http://creativecommons.org/licenses/by/4.0/ open access Atribución 4.0 Internacional Springer Nature