Bridging length scales from molecules to the whole organism by cryoCLEM and cryoET. (12th August 2022)

• Record Type: Journal Article
• Authors: Lovatt, Megan; Leistner, Conny; Frank, René A. W.
• Is Part Of: Faraday discussions. Volume 240(2022)
• Page Start: 114
• Page End: 126
• Publication Date: 2022-08-12
• Subjects: Chemistry -- Periodicals; Metallurgy -- Periodicals; Electrochemistry -- Periodicals; 540
• DOI: 10.1039/d2fd00081d
• Languages: English
• ISSNs: 1359-6640

Abstract

Abstract : Experimental workflows combining mouse genetics, cryogenic correlated light and electron microscopy and cryo-electron tomography to bridge length scales from the whole organism to molecules. Abstract : Resolving atomic structures of isolated proteins has uncovered mechanisms and fundamental processes in biology. However, many functions can only be tested in the context of intact cells and tissues that are many orders of magnitude larger than the macromolecules on which they depend. Therefore, methods that interrogate macromolecular structure in situ provide a means of directly relating structure to function across length scales. Here, we developed several workflows using cryogenic correlated light and electron microscopy (cryoCLEM) and electron tomography (cryoET) that can bridge this gap to reveal the molecular infrastructure that underlies higher order functions within cells and tissues. We also describe experimental design considerations, including cryoCLEM labelling, sample preparation, and quality control, for determining the in situ molecular architectures within native, hydrated cells and tissues.