TY - JOUR T1 - Precision super-resolution cryo-correlative light and electron microscopy for rapid <em>in situ</em> structural analyses of optogenetically-positioned organelles JF - bioRxiv DO - 10.1101/2022.11.22.516823 SP - 2022.11.22.516823 AU - G.M.I. Redpath AU - J. Rae AU - Y. Yao AU - J. Ruan AU - M.L. Cagigas AU - R. Whan AU - E.C. Hardeman AU - P.W. Gunning AU - V. Ananthanarayanan AU - R.G. Parton AU - N.A. Ariotti Y1 - 2022/01/01 UR - http://biorxiv.org/content/early/2022/11/23/2022.11.22.516823.abstract N2 - Unambiguous targeting of cellular structures for in situ cryo-electron microscopy in the heterogeneous, dense, and compacted environment of the cytoplasm remains challenging. Here we have developed a novel cryogenic correlative light and electron microscopy (cryo-CLEM) workflow which combines thin cells grown on a mechanically defined substratum to rapidly analyse organelles and macromolecular complexes in the cell by cryo-electron tomography (cryo-ET). We coupled these advancements with optogenetics to redistribute perinuclear-localised organelles to the cell periphery for cryo-ET. This reliable and robust workflow allows for fast in situ analyses without the requirement for cryo-focused ion beam milling. We have developed a protocol where cells can be frozen, imaged by cryo-fluorescence microscopy and ready for batch cryo-ET within a day.Competing Interest StatementThe authors have declared no competing interest. ER -