ClearT immersion optical clearing method for intact 3D spheroids imaging through confocal laser scanning microscopy
Main Author: | |
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Publication Date: | 2018 |
Other Authors: | , , |
Format: | Article |
Language: | eng |
Source: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Download full: | http://hdl.handle.net/10400.6/4735 |
Summary: | Spheroids 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. |
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ClearT immersion optical clearing method for intact 3D spheroids imaging through confocal laser scanning microscopyClearTSpheroidsPropidium iodideFluorescenceConfocal microscopySpheroids 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.ElsevieruBibliorumCosta, Elisabete C.Moreira, AndréDiogo, Duarte Miguel de MeloCorreia, Ilídio Joaquim Sobreira2018-04-09T11:27:34Z20182018-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.6/4735engCosta, 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.https://doi.org/10.1016/j.optlastec.2018.04.002metadata only accessinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-12-15T09:42:03Zoai:ubibliorum.ubi.pt:10400.6/4735Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T00:45:46.815959Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
ClearT immersion optical clearing method for intact 3D spheroids imaging through confocal laser scanning microscopy |
title |
ClearT immersion optical clearing method for intact 3D spheroids imaging through confocal laser scanning microscopy |
spellingShingle |
ClearT immersion optical clearing method for intact 3D spheroids imaging through confocal laser scanning microscopy Costa, Elisabete C. ClearT Spheroids Propidium iodide Fluorescence Confocal microscopy |
title_short |
ClearT immersion optical clearing method for intact 3D spheroids imaging through confocal laser scanning microscopy |
title_full |
ClearT immersion optical clearing method for intact 3D spheroids imaging through confocal laser scanning microscopy |
title_fullStr |
ClearT immersion optical clearing method for intact 3D spheroids imaging through confocal laser scanning microscopy |
title_full_unstemmed |
ClearT immersion optical clearing method for intact 3D spheroids imaging through confocal laser scanning microscopy |
title_sort |
ClearT immersion optical clearing method for intact 3D spheroids imaging through confocal laser scanning microscopy |
author |
Costa, Elisabete C. |
author_facet |
Costa, Elisabete C. Moreira, André Diogo, Duarte Miguel de Melo Correia, Ilídio Joaquim Sobreira |
author_role |
author |
author2 |
Moreira, André Diogo, Duarte Miguel de Melo Correia, Ilídio Joaquim Sobreira |
author2_role |
author author author |
dc.contributor.none.fl_str_mv |
uBibliorum |
dc.contributor.author.fl_str_mv |
Costa, Elisabete C. Moreira, André Diogo, Duarte Miguel de Melo Correia, Ilídio Joaquim Sobreira |
dc.subject.por.fl_str_mv |
ClearT Spheroids Propidium iodide Fluorescence Confocal microscopy |
topic |
ClearT Spheroids Propidium iodide Fluorescence Confocal microscopy |
description |
Spheroids 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. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-04-09T11:27:34Z 2018 2018-01-01T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10400.6/4735 |
url |
http://hdl.handle.net/10400.6/4735 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Costa, 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. https://doi.org/10.1016/j.optlastec.2018.04.002 |
dc.rights.driver.fl_str_mv |
metadata only access info:eu-repo/semantics/openAccess |
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metadata only access |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Elsevier |
publisher.none.fl_str_mv |
Elsevier |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799136355395043328 |