Indirect correlative light and electron microscopy (iCLEM) : a novel pipeline for multiscale quantification of structure from molecules to organs
- Author
- Heather L Struckman, Nicolae Moise, Bieke Vanslembrouck, Nathan Rothacker, Zhenhui Chen, Jolanda van Hengel (UGent) , Seth H Weinberg and Rengasayee Veeraraghavan
- Organization
- Abstract
- 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.
- Keywords
- multiscale imagingquantitative image analysis, high throughput, correlative light and electron microscopy (CLEM), computational modeling
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Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-01HWM9VKHBZKYEFHG3SMQB1T70
- MLA
- Struckman, Heather L., et al. “Indirect Correlative Light and Electron Microscopy (ICLEM) : A Novel Pipeline for Multiscale Quantification of Structure from Molecules to Organs.” MICROSCOPY AND MICROANALYSIS, vol. 30, no. 2, 2024, pp. 318–33, doi:10.1093/mam/ozae021.
- APA
- Struckman, H. L., Moise, N., Vanslembrouck, B., Rothacker, N., Chen, Z., van Hengel, J., … Veeraraghavan, R. (2024). Indirect correlative light and electron microscopy (iCLEM) : a novel pipeline for multiscale quantification of structure from molecules to organs. MICROSCOPY AND MICROANALYSIS, 30(2), 318–333. https://doi.org/10.1093/mam/ozae021
- Chicago author-date
- Struckman, Heather L, Nicolae Moise, Bieke Vanslembrouck, Nathan Rothacker, Zhenhui Chen, Jolanda van Hengel, Seth H Weinberg, and Rengasayee Veeraraghavan. 2024. “Indirect Correlative Light and Electron Microscopy (ICLEM) : A Novel Pipeline for Multiscale Quantification of Structure from Molecules to Organs.” MICROSCOPY AND MICROANALYSIS 30 (2): 318–33. https://doi.org/10.1093/mam/ozae021.
- Chicago author-date (all authors)
- Struckman, Heather L, Nicolae Moise, Bieke Vanslembrouck, Nathan Rothacker, Zhenhui Chen, Jolanda van Hengel, Seth H Weinberg, and Rengasayee Veeraraghavan. 2024. “Indirect Correlative Light and Electron Microscopy (ICLEM) : A Novel Pipeline for Multiscale Quantification of Structure from Molecules to Organs.” MICROSCOPY AND MICROANALYSIS 30 (2): 318–333. doi:10.1093/mam/ozae021.
- Vancouver
- 1.Struckman HL, Moise N, Vanslembrouck B, Rothacker N, Chen Z, van Hengel J, et al. Indirect correlative light and electron microscopy (iCLEM) : a novel pipeline for multiscale quantification of structure from molecules to organs. MICROSCOPY AND MICROANALYSIS. 2024;30(2):318–33.
- IEEE
- [1]H. L. Struckman et al., “Indirect correlative light and electron microscopy (iCLEM) : a novel pipeline for multiscale quantification of structure from molecules to organs,” MICROSCOPY AND MICROANALYSIS, vol. 30, no. 2, pp. 318–333, 2024.
@article{01HWM9VKHBZKYEFHG3SMQB1T70,
abstract = {{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.}},
author = {{Struckman, Heather L and Moise, Nicolae and Vanslembrouck, Bieke and Rothacker, Nathan and Chen, Zhenhui and van Hengel, Jolanda and Weinberg, Seth H and Veeraraghavan, Rengasayee}},
issn = {{1431-9276}},
journal = {{MICROSCOPY AND MICROANALYSIS}},
keywords = {{multiscale imagingquantitative image analysis,high throughput,correlative light and electron microscopy (CLEM),computational modeling}},
language = {{eng}},
number = {{2}},
pages = {{318--333}},
title = {{Indirect correlative light and electron microscopy (iCLEM) : a novel pipeline for multiscale quantification of structure from molecules to organs}},
url = {{http://doi.org/10.1093/mam/ozae021}},
volume = {{30}},
year = {{2024}},
}
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