Correlative confocal and scanning electron microscopy of cultured cells without using dedicated equipment

This protocol enables correlative light and electron microscopy (CLEM) imaging of cell surface features without using dedicated equipment. Cells are cultured and fixed on transparent substrates for confocal microscopy imaging. No conductive coating is employed in the scanning electron microscopy workflow, providing a clean cell surface observation, with fiducial markers assisting alignment of optical and topographical images. This protocol describes CLEM imaging for midbody remnants in MDCK cells but can also be applied to different cell types and surface features. For complete details on the use and execution of this protocol, please refer to Casares-Arias et al. (2020). https://doi.org/10.1016/j.isci.2020.101244The expert technical advice of the Optical and Confocal Microscopy Facility of CBMSO is gratefully acknowledged. We acknowledge the Micro and Nanofabrication Laboratory of the Instituto de Micro y Nanotecnología (MiNa), which is funded by the Comunidad de Madrid (S2018/NMT-4291 TEC2-SPACE), MICIN (project CSIC13-4E-1794), and EU (FEDER, FSE), for support with SEM infrastructure. Research in the laboratory of MAA was supported by a grant (PGC2018-095643-B-I00) from the Spanish Ministerio de Ciencia e Innovación (MICINN), Agencia Estatal de Investigación, and the Fondo Europeo de Desarrollo Regional, European Union (MICINN/AEI/FEDER, EU). A contract (FPU14/00295) and a short-term fellowship from EMBO to J.C.A. are also acknowledged.Peer reviewe