A quantitative metric for the comparative evaluation of optical clearing protocols for 3D multicellular spheroids

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dc.contributor.author Diosdi, Akos
dc.contributor.author Hirling, Dominik
dc.contributor.author Kovacs, Maria
dc.contributor.author Toth, Timea
dc.contributor.author Harmati, Maria
dc.contributor.author Koos, Krisztian
dc.contributor.author Buzas, Krisztina
dc.contributor.author Piccinini, Filippo
dc.contributor.author Horvath, Peter
dc.date.accessioned 2021-10-07T08:14:01Z
dc.date.available 2021-10-07T08:14:01Z
dc.date.issued 2021
dc.identifier.citation Diosdi , A , Hirling , D , Kovacs , M , Toth , T , Harmati , M , Koos , K , Buzas , K , Piccinini , F & Horvath , P 2021 , ' A quantitative metric for the comparative evaluation of optical clearing protocols for 3D multicellular spheroids ' , Computational and Structural Biotechnology Journal , vol. 19 , pp. 1233-1243 . https://doi.org/10.1016/j.csbj.2021.01.040
dc.identifier.other PURE: 169235582
dc.identifier.other PURE UUID: 53fc0117-ca75-44eb-af7b-b2111bc1d829
dc.identifier.other WOS: 000684854500017
dc.identifier.uri http://hdl.handle.net/10138/335043
dc.description.abstract 3D multicellular spheroids quickly emerged as in vitro models because they represent the in vivo tumor environment better than standard 2D cell cultures. However, with current microscopy technologies, it is difficult to visualize individual cells in the deeper layers of 3D samples mainly because of limited light penetration and scattering. To overcome this problem several optical clearing methods have been proposed but defining the most appropriate clearing approach is an open issue due to the lack of a gold standard metric. Here, we propose a guideline for 3D light microscopy imaging to achieve single-cell resolution. The guideline includes a validation experiment focusing on five optical clearing protocols. We review and compare seven quality metrics which quantitatively characterize the imaging quality of spheroids. As a test environment, we have created and shared a large 3D dataset including approximately hundred fluorescently stained and optically cleared spheroids. Based on the results we introduce the use of a novel quality metric as a promising method to serve as a gold standard, applicable to compare optical clearing protocols, and decide on the most suitable one for a particular experiment. (C) 2021 The Authors. Published by Elsevier B.V. on behalf of Research Network of Computational and Structural Biotechnology. en
dc.format.extent 11
dc.language.iso eng
dc.relation.ispartof Computational and Structural Biotechnology Journal
dc.rights cc_by
dc.rights.uri info:eu-repo/semantics/openAccess
dc.subject Optical tissue clearing
dc.subject Spheroid
dc.subject Light-sheet microscopy
dc.subject Focus metrics
dc.subject CELL
dc.subject MICROSCOPY
dc.subject CLEAR(T)
dc.subject 1182 Biochemistry, cell and molecular biology
dc.title A quantitative metric for the comparative evaluation of optical clearing protocols for 3D multicellular spheroids en
dc.type Article
dc.contributor.organization Institute for Molecular Medicine Finland
dc.description.reviewstatus Peer reviewed
dc.relation.doi https://doi.org/10.1016/j.csbj.2021.01.040
dc.relation.issn 2001-0370
dc.rights.accesslevel openAccess
dc.type.version publishedVersion

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