Indirect Correlative Light and Electron Microscopy (iCLEM): A Novel Pipeline for Multiscale Quantification of Structure From Molecules to Organs.

Autor:	Struckman HL; Department of Biomedical Engineering, College of Engineering, 2124 Fontana Labs, 140 W. 19th Ave, The Ohio State University, Columbus, OH 43210, USA.; The Frick Center for Heart Failure and Arrhythmia, Dorothy M. Davis Heart and Lung Research Institute, College of Medicine, The Ohio State University Wexner Medical Center, 2255 Kenny Rd, Rm 5189, Pelotonia Research Center, Columbus, OH 43210, USA., Moise N; The Frick Center for Heart Failure and Arrhythmia, Dorothy M. Davis Heart and Lung Research Institute, College of Medicine, The Ohio State University Wexner Medical Center, 2255 Kenny Rd, Rm 5189, Pelotonia Research Center, Columbus, OH 43210, USA., Vanslembrouck B; Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Rd, Berkeley, CA 94720, USA.; Medical Cell Biology Research Group, Department of Human Structure and Repair, Faculty of Medicine and Health Sciences, Ghent University, Corneel Heymanslaan 10, Building B, Entrance 36, 9000 Ghent, Belgium., Rothacker N; Department of Biomedical Engineering, College of Engineering, 2124 Fontana Labs, 140 W. 19th Ave, The Ohio State University, Columbus, OH 43210, USA.; The Frick Center for Heart Failure and Arrhythmia, Dorothy M. Davis Heart and Lung Research Institute, College of Medicine, The Ohio State University Wexner Medical Center, 2255 Kenny Rd, Rm 5189, Pelotonia Research Center, Columbus, OH 43210, USA., Chen Z; Krannert Cardiovascular Research Center, Department of Medicine, Indiana University, Room E400, 1801 N. Senate Blvd., Suite E400, Indianapolis, IN 46202, USA., van Hengel J; Medical Cell Biology Research Group, Department of Human Structure and Repair, Faculty of Medicine and Health Sciences, Ghent University, Corneel Heymanslaan 10, Building B, Entrance 36, 9000 Ghent, Belgium., Weinberg SH; Department of Biomedical Engineering, College of Engineering, 2124 Fontana Labs, 140 W. 19th Ave, The Ohio State University, Columbus, OH 43210, USA.; The Frick Center for Heart Failure and Arrhythmia, Dorothy M. Davis Heart and Lung Research Institute, College of Medicine, The Ohio State University Wexner Medical Center, 2255 Kenny Rd, Rm 5189, Pelotonia Research Center, Columbus, OH 43210, USA., Veeraraghavan R; Department of Biomedical Engineering, College of Engineering, 2124 Fontana Labs, 140 W. 19th Ave, The Ohio State University, Columbus, OH 43210, USA.; The Frick Center for Heart Failure and Arrhythmia, Dorothy M. Davis Heart and Lung Research Institute, College of Medicine, The Ohio State University Wexner Medical Center, 2255 Kenny Rd, Rm 5189, Pelotonia Research Center, Columbus, OH 43210, USA.
Jazyk:	angličtina
Zdroj:	Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada [Microsc Microanal] 2024 Apr 29; Vol. 30 (2), pp. 318-333.
DOI:	10.1093/mam/ozae021
Abstrakt:	Correlative light and electron microscopy (CLEM) methods are powerful methods that combine molecular organization (from light microscopy) with ultrastructure (from electron microscopy). However, CLEM methods pose high cost/difficulty barriers to entry and have very low experimental throughput. Therefore, we have developed an indirect correlative light and electron microscopy (iCLEM) pipeline to sidestep the rate-limiting steps of CLEM (i.e., preparing and imaging the same samples on multiple microscopes) and correlate multiscale structural data gleaned from separate samples imaged using different modalities by exploiting biological structures identifiable by both light and electron microscopy as intrinsic fiducials. We demonstrate here an application of iCLEM, where we utilized gap junctions and mechanical junctions between muscle cells in the heart as intrinsic fiducials to correlate ultrastructural measurements from transmission electron microscopy (TEM), and focused ion beam scanning electron microscopy (FIB-SEM) with molecular organization from confocal microscopy and single molecule localization microscopy (SMLM). We further demonstrate how iCLEM can be integrated with computational modeling to discover structure-function relationships. Thus, we present iCLEM as a novel approach that complements existing CLEM methods and provides a generalizable framework that can be applied to any set of imaging modalities, provided suitable intrinsic fiducials can be identified. Competing Interests: Conflict of Interest: The authors declare none. (© The Author(s) 2024. Published by Oxford University Press on behalf of the Microscopy Society of America. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)
Databáze:	MEDLINE
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