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Beyond the whole-mount phenotype: high-resolution imaging in fluorescence-based applications on zebrafish

2019Artigo científico Acesso aberto
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dc.contributor.authorOralová, Veronika
dc.contributor.authorRosa, Joana
dc.contributor.authorSoenens, Mieke
dc.contributor.authorBek, Jan Willem
dc.contributor.authorWillaert, Andy
dc.contributor.authorWitten, Paul Eckhard
dc.contributor.authorHuysseune, Ann
dc.date.accessioned2020-08-03T12:48:46Z
dc.date.available2020-08-03T12:48:46Z
dc.date.issued2019
dc.description.abstractZebrafish is now widely used in biomedical research as a model for human diseases, but the relevance of the model depends on a rigorous analysis of the phenotypes obtained. Many zebrafish disease models, experimental techniques and manipulations take advantage of fluorescent reporter molecules. However, phenotypic analysis often does not go beyond establishing overall distribution patterns of the fluorophore in whole-mount embryos or using vibratome or paraffin sections with poor preservation of tissue architecture and limited resolution. Obtaining high-resolution data of fluorescent signals at the cellular level from internal structures mostly depends on the availability of expensive imaging technology. Here, we propose a new and easily applicable protocol for embedding and sectioning of zebrafish embryos using in-house prepared glycol methacrylate (GMA) plastic that is suited for preservation of fluorescent signals (including photoactivatable fluorophores) without the need for antibodies. Four main approaches are described, all involving imaging fluorescent signals on semithin (3 µm or less) sections. These include sectioning transgenic animals, whole-mount immunostained embryos, cell tracking, as well as on-section enzyme histochemistry.pt_PT
dc.description.sponsorshipAgência financiadora Ghent University BOF24J2015001401pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.1242/bio.042374pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.1/14572
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherCompany of Biologistspt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectZebrafishpt_PT
dc.subjectGMApt_PT
dc.subjectGFPpt_PT
dc.subjectFluorophorespt_PT
dc.subjectImmunofluorescencept_PT
dc.subjectCell trackingpt_PT
dc.subjectTRAPpt_PT
dc.titleBeyond the whole-mount phenotype: high-resolution imaging in fluorescence-based applications on zebrafishpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.issue5pt_PT
oaire.citation.startPagebio042374pt_PT
oaire.citation.titleBiology Openpt_PT
oaire.citation.volume8pt_PT
person.familyNameRosa
person.givenNameJoana
person.identifier.ciencia-id0B18-8AEC-875C
person.identifier.orcid0000-0001-7947-2681
person.identifier.scopus-author-id36505067900
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublication181f1017-5086-40dc-9edc-a289aaa2d263
relation.isAuthorOfPublication.latestForDiscovery181f1017-5086-40dc-9edc-a289aaa2d263

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