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ClearT immersion optical clearing method for intact 3D spheroids imaging through confocal laser scanning microscopy

2018Journal article Restricted access
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dc.contributor.authorCosta, Elisabete C.
dc.contributor.authorMoreira, André
dc.contributor.authorDiogo, Duarte Miguel de Melo
dc.contributor.authorCorreia, Ilídio Joaquim Sobreira
dc.date.accessioned2018-04-09T11:27:34Z
dc.date.available2018-04-09T11:27:34Z
dc.date.issued2018
dc.description.abstractSpheroids are 3D in vitro platforms that fill the gap between the 2D cell cultures and animal models on the therapeutics development pipeline. Yet, the methods and equipment used in the in vitro assays are optimized for the analysis of cells cultured as monolayers. For instance, confocal laser scanning microscopy (CLSM) does not allow the observation of thick intact spheroids due to light penetration issues. To overcome this limitation, spheroids treatment with clearing agents started to be explored. Herein, we demonstrate for the first time the application of ClearT clearing method for the imaging of propidium iodide (PI) stained spheroids by CLSM. The results demonstrate that the ClearT is a reversible clearing method that does not influence the structure of the spheroid and significantly improved the PI signal penetration depth in about 43%. Additionally, ClearT also enhanced the cells imaging within the spheroid by increasing the cross-penetration depth in 46.6% at 100 µm of depth. Overall, the results show that ClearT method may allow the improvement of the CLSM accuracy on the evaluation of the cellular death within spheroids prompted by therapeutics.pt_PT
dc.description.versioninfo:eu-repo/semantics/acceptedVersionpt_PT
dc.identifier.citationCosta, E.C., Moreira, A.F., de Melo-Diogo, Correia, I.J. (2018) "ClearT immersion optical clearing method for intact 3D spheroids imaging through confocal laser scanning microscopy”, Optics and Laser Technology, accepted for publication.pt_PT
dc.identifier.doihttps://doi.org/10.1016/j.optlastec.2018.04.002
dc.identifier.urihttp://hdl.handle.net/10400.6/4735
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherElsevierpt_PT
dc.relationDevelopment of layer-by-layer polymeric microneedles for localized delivery of therapeutics to cervical cancer
dc.relationMULTIFUNCTIONAL GRAPHENE-OXIDE NANOCARRIERS FOR DRUG DELIVERY AND PHOTOTHERMAL THERAPY OF LUNG CANCER
dc.relation3D multicellular spheroids as high throughput platforms to screen novel combinatory therapies for treatment of pancreatic cancer
dc.subjectClearTpt_PT
dc.subjectSpheroidspt_PT
dc.subjectPropidium iodidept_PT
dc.subjectFluorescencept_PT
dc.subjectConfocal microscopypt_PT
dc.titleClearT immersion optical clearing method for intact 3D spheroids imaging through confocal laser scanning microscopypt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleDevelopment of layer-by-layer polymeric microneedles for localized delivery of therapeutics to cervical cancer
oaire.awardTitleMULTIFUNCTIONAL GRAPHENE-OXIDE NANOCARRIERS FOR DRUG DELIVERY AND PHOTOTHERMAL THERAPY OF LUNG CANCER
oaire.awardTitle3D multicellular spheroids as high throughput platforms to screen novel combinatory therapies for treatment of pancreatic cancer
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/5876/UID%2FMulti%2F00709%2F2013/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/POR_CENTRO/SFRH%2FBD%2F109482%2F2015/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//SFRH%2FBD%2F103506%2F2014/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//SFRH%2FBD%2F103507%2F2014/PT
oaire.citation.endPage99
oaire.citation.startPage94
oaire.citation.titleOptics and Laser Technologypt_PT
oaire.fundingStream5876
oaire.fundingStreamPOR_CENTRO
person.familyNameda Rocha Costa
person.familyNameMoreira
person.familyNameMiguel de Melo Diogo
person.familyNameJoaquim Sobreira Correia
person.givenNameElisabete Cristina
person.givenNameAndré Ferreira
person.givenNameDuarte
person.givenNameIlídio
person.identifier1161643
person.identifierC2z5x04AAAAJ
person.identifierUMbJ1KMAAAAJ
person.identifier.ciencia-idB314-4CF0-6633
person.identifier.ciencia-id4C1E-012B-83CE
person.identifier.ciencia-id9C10-9BD8-634E
person.identifier.ciencia-idF610-7373-DC81
person.identifier.orcid0000-0002-0490-0095
person.identifier.orcid0000-0002-0604-2506
person.identifier.orcid0000-0001-9984-3603
person.identifier.orcid0000-0003-1613-9675
person.identifier.scopus-author-id55805850100
person.identifier.scopus-author-id56401521300
person.identifier.scopus-author-id56266511300
person.identifier.scopus-author-id7003557499
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.embargofctCopyright cedido à editora no momento da publicaçãopt_PT
rcaap.rightsclosedAccesspt_PT
rcaap.typearticlept_PT
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