Application of ethyl cinnamate based optical tissue clearing and expansion microscopy combined with retrograde perfusion for 3D lung imaging

• Verfasst von: Sun, Quanchao [VerfasserIn]
• Verfasst von: Picascia, Tiziana [VerfasserIn]
• Verfasst von: Khan, Arif ul Maula [VerfasserIn]
• Verfasst von: Brenna, Cinzia [VerfasserIn]
• Verfasst von: Heuveline, Vincent [VerfasserIn]
• Verfasst von: Schmaus, Anja [VerfasserIn]
• Verfasst von: Sleeman, Jonathan P. [VerfasserIn]
• Verfasst von: Gretz, Norbert [VerfasserIn]
• Titel: Application of ethyl cinnamate based optical tissue clearing and expansion microscopy combined with retrograde perfusion for 3D lung imaging
• Verf.angabe: Quanchao Sun, Tiziana Picascia, Arif ul Maula Khan, Cinzia Brenna, Vincent Heuveline, Anja Schmaus, Jonathan P. Sleeman and Norbert Gretz
• E-Jahr: 2020
• Jahr: 12 Oct 2020
• Umfang: 16 S.
• Fussnoten: Gesehen am 27.01.2025
• Titel Quelle: Enthalten in: Experimental lung research
• Ort Quelle: Abingdon : Taylor & Francis Group, 1980
• Jahr Quelle: 2020
• Band/Heft Quelle: 46(2020), 10, Seite 393-408
• ISSN Quelle: 1521-0499
• DOI: doi:10.1080/01902148.2020.1829183
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: Ethyl cinnamate
• Sach-SW: expansion microscopy
• Sach-SW: lung
• Sach-SW: retrograde perfusion
• Sach-SW: three-dimensional imaging
• K10plus-PPN: 191567896X

Abstract

3 D imaging of the lung is not a trivial undertaking as during preparation the lung may collapse. Also serial sections and scans followed by 3 D reconstruction may lead to artifacts. The present study aims to figure out the best way to perform 3 D imaging in lung research. We applied an optical tissue clearing (OTC) method, which uses ethyl cinnamate (ECi) as a fast, nontoxic and cheap clearing solvent, for 3 D imaging of retrograde perfused lungs by laser confocal fluorescence microscopy and light sheet fluorescence microscopy. We also introduced expansion microscopy (ExM), a cutting-edge technique, in 3 D imaging of lungs. We examined and compared the usefulness of these techniques for 3 D lung imaging. The ExM protocol was further extended to paraffin-embedded lung metastases blocks. The MHI148-PEI labeled lung vasculature was visualized by retrograde perfusion combined with trachea ligation and ECi based OTC. As compared with trans-cardiac perfusion, the retrograde perfusion results in a better maintenance of lung morphology. 3 D structure of alveoli, vascular branches and cilia in lung were revealed by immunofluorescence staining after ExM. 3 D distribution of microvasculature and neutrophil cells in 10 years old paraffin-embedded lung metastases were analyzed by ExM. The retrograde perfusion combined with trachea ligation technique could be applied in the lung research in mice. 3 D structure of lung vasculature can be visualized by MHI148-PEI perfusion and ECi based OTC in an efficient way. ExM and immunofluorescence staining protocol is highly recommended to perform 3 D imaging of fresh fixed lung as well as paraffin-embedded lung blocks.