A Cryo‐to‐Liquid Phase Correlative Light Electron Microscopy Workflow for the Visualization of Biological Processes in Graphene Liquid Cells
Advanced Functional Materials, vol. 35
Abstract
Abstract Liquid phase electron microscopy (LP‐EM) has emerged as a powerful technique for in situ observation of material formation in liquid. Especially the use of graphene as window material provides new opportunities to image biological processes because of graphene's molecular thickness and electron scavenger capabilities. However, in most cases the process of interest is initiated when the graphene liquid cells (GLCs) are sealed, meaning that the process cannot be imaged at early timepoints. Here, a novel cryogenic/liquid phase correlative light/electron microscopy workflow that addresses the delay time between graphene encapsulation and the start of the imaging, while combining the advantages of fluorescence and electron microscopy is reported. This workflow allows imaging to be initiated at a predetermined space and time by vitrifying and thawing at a selected time point. The workflow is demonstrated first by observing multiple day crystallization processes and subsequently highlight its potential by observing a biological process: the complexation of calciprotein particles. The ability to correlate the dynamic complexation observed in a GLC with cryogenic TEM and dynamic light scattering, confirms the validity of observations and underlines the exciting possibilities for LP‐EM in biology.
Authors 10
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Radboud University Nijmegen · Radboud University Medical Center
Affiliation as printed
Department of Medical BioSciences Research Institute for Medical Innovations Radboud University Medical Center Geert Grooteplein Zuid 28 Nijmegen 6525 GA Netherlands
Electron Microscopy Center Radboudumc Technology Center Microscopy Radboud University Medical Center Geert Grooteplein Noord 29 Nijmegen EZ 6525 Netherlands
Department of Medical BioSciences, Research Institute for Medical Innovations, Radboud University Medical Center, Geert Grooteplein Zuid 28, Nijmegen, 6525 GA Netherlands
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Radboud University Nijmegen · Radboud University Medical Center
Affiliation as printed
Department of Medical BioSciences Research Institute for Medical Innovations Radboud University Medical Center Geert Grooteplein Zuid 28 Nijmegen 6525 GA Netherlands
Electron Microscopy Center Radboudumc Technology Center Microscopy Radboud University Medical Center Geert Grooteplein Noord 29 Nijmegen EZ 6525 Netherlands
Department of Medical BioSciences, Research Institute for Medical Innovations, Radboud University Medical Center, Geert Grooteplein Zuid 28, Nijmegen, 6525 GA Netherlands
Electron Microscopy Center, Radboudumc Technology Center Microscopy, Radboud University Medical Center, Geert Grooteplein Noord 29, Nijmegen, EZ 6525 Netherlands
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Radboud University Nijmegen · Radboud University Medical Center
Affiliation as printed
Department of Medical BioSciences Research Institute for Medical Innovations Radboud University Medical Center Geert Grooteplein Zuid 28 Nijmegen 6525 GA Netherlands
Electron Microscopy Center Radboudumc Technology Center Microscopy Radboud University Medical Center Geert Grooteplein Noord 29 Nijmegen EZ 6525 Netherlands
Department of Medical BioSciences, Research Institute for Medical Innovations, Radboud University Medical Center, Geert Grooteplein Zuid 28, Nijmegen, 6525 GA Netherlands
Electron Microscopy Center, Radboudumc Technology Center Microscopy, Radboud University Medical Center, Geert Grooteplein Noord 29, Nijmegen, EZ 6525 Netherlands
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Radboud University Nijmegen · Radboud University Medical Center
Affiliation as printed
Department of Medical BioSciences Research Institute for Medical Innovations Radboud University Medical Center Geert Grooteplein Zuid 28 Nijmegen 6525 GA Netherlands
Electron Microscopy Center Radboudumc Technology Center Microscopy Radboud University Medical Center Geert Grooteplein Noord 29 Nijmegen EZ 6525 Netherlands
Department of Medical BioSciences, Research Institute for Medical Innovations, Radboud University Medical Center, Geert Grooteplein Zuid 28, Nijmegen, 6525 GA Netherlands
Electron Microscopy Center, Radboudumc Technology Center Microscopy, Radboud University Medical Center, Geert Grooteplein Noord 29, Nijmegen, EZ 6525 Netherlands
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Radboud University Nijmegen · Radboud University Medical Center
Affiliation as printed
Department of Medical BioSciences Research Institute for Medical Innovations Radboud University Medical Center Geert Grooteplein Zuid 28 Nijmegen 6525 GA Netherlands
Electron Microscopy Center Radboudumc Technology Center Microscopy Radboud University Medical Center Geert Grooteplein Noord 29 Nijmegen EZ 6525 Netherlands
Department of Medical BioSciences, Research Institute for Medical Innovations, Radboud University Medical Center, Geert Grooteplein Zuid 28, Nijmegen, 6525 GA Netherlands
Electron Microscopy Center, Radboudumc Technology Center Microscopy, Radboud University Medical Center, Geert Grooteplein Noord 29, Nijmegen, EZ 6525 Netherlands
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Universitätsklinikum Aachen · RWTH Aachen University
Affiliation as printed
Helmholtz Institute for Biomedical Engineering Biointerface Laboratory RWTH Aachen University Hospital Pauwelsstrasse 30 52074 Aachen Germany
Helmholtz Institute for Biomedical Engineering, Biointerface Laboratory RWTH Aachen University Hospital, Pauwelsstrasse 30, 52074 Aachen, Germany
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Universitätsklinikum Aachen · RWTH Aachen University
Affiliation as printed
Helmholtz Institute for Biomedical Engineering Biointerface Laboratory RWTH Aachen University Hospital Pauwelsstrasse 30 52074 Aachen Germany
Helmholtz Institute for Biomedical Engineering, Biointerface Laboratory RWTH Aachen University Hospital, Pauwelsstrasse 30, 52074 Aachen, Germany
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Radboud University Nijmegen · Radboud University Medical Center
Affiliation as printed
Department of Medical BioSciences Research Institute for Medical Innovations Radboud University Medical Center Geert Grooteplein Zuid 28 Nijmegen 6525 GA Netherlands
Electron Microscopy Center Radboudumc Technology Center Microscopy Radboud University Medical Center Geert Grooteplein Noord 29 Nijmegen EZ 6525 Netherlands
Department of Medical BioSciences, Research Institute for Medical Innovations, Radboud University Medical Center, Geert Grooteplein Zuid 28, Nijmegen, 6525 GA Netherlands
Electron Microscopy Center, Radboudumc Technology Center Microscopy, Radboud University Medical Center, Geert Grooteplein Noord 29, Nijmegen, EZ 6525 Netherlands
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Elena Macías‐Sánchez corresponding
Radboud University Nijmegen · Universidad de Granada · Radboud University Medical Center
Affiliation as printed
Department of Medical BioSciences Research Institute for Medical Innovations Radboud University Medical Center Geert Grooteplein Zuid 28 Nijmegen 6525 GA Netherlands
Department of Stratigraphy and Palaeontolgy University of Granada Avenida Fuente Nueva S/N CP Granada 18071 Spain
Electron Microscopy Center Radboudumc Technology Center Microscopy Radboud University Medical Center Geert Grooteplein Noord 29 Nijmegen EZ 6525 Netherlands
Department of Medical BioSciences, Research Institute for Medical Innovations, Radboud University Medical Center, Geert Grooteplein Zuid 28, Nijmegen, 6525 GA Netherlands
Department of Stratigraphy and Palaeontolgy, University of Granada, Avenida Fuente Nueva S/N CP, Granada, 18071 Spain
Electron Microscopy Center, Radboudumc Technology Center Microscopy, Radboud University Medical Center, Geert Grooteplein Noord 29, Nijmegen, EZ 6525 Netherlands
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Nico A. J. M. Sommerdijk corresponding
Radboud University Nijmegen · Radboud University Medical Center
Affiliation as printed
Department of Medical BioSciences Research Institute for Medical Innovations Radboud University Medical Center Geert Grooteplein Zuid 28 Nijmegen 6525 GA Netherlands
Electron Microscopy Center Radboudumc Technology Center Microscopy Radboud University Medical Center Geert Grooteplein Noord 29 Nijmegen EZ 6525 Netherlands
Department of Medical BioSciences, Research Institute for Medical Innovations, Radboud University Medical Center, Geert Grooteplein Zuid 28, Nijmegen, 6525 GA Netherlands
Electron Microscopy Center, Radboudumc Technology Center Microscopy, Radboud University Medical Center, Geert Grooteplein Noord 29, Nijmegen, EZ 6525 Netherlands
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