Quantitative 3D Imaging of Trypanosoma cruzi-Infected Cells, Dormant Amastigotes, and T Cells in Intact Clarified Organs

Autores
Sánchez Valdéz, Fernando Javier; Padilla, Angel Marcelo; Bustamante, Juan Manuel; Hawkins, Caleb W. D.; Tarleton, Rick L.
Año de publicación
2021
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Reliable detection of Trypanosoma cruzi (T. cruzi) in vivo infections have long been needed to understand the complex biology of Chagas disease and to accurately evaluate the outcome of treatment regimens. Here, an integrated pipeline for automated quantification of T. cruzi-infected cells in 3D-reconstructed, cleared organs was developed. Light-sheet fluorescent microscopy allows us to accurately visualize and quantify not only actively proliferating but also dormant T. cruzi parasites and immune effector cells in whole-organs or tissues. Also, CUBIC-HistoVision pipeline to obtain uniform labeling of cleared organs with antibodies and nuclear stains was successfully adopted. Tissue clearing coupled to 3D immunostaining provides an unbiased approach to comprehensively evaluate drug treatment protocols, improve the understanding of the cellular organization of T. cruzi-infected tissues and is expected to advance discoveries related to anti-T. cruzi immune responses, tissue damage and repair in Chagas disease.
Fil: Sánchez Valdéz, Fernando Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina
Fil: Padilla, Angel Marcelo. University of Georgia; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Bustamante, Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. University of Georgia; Estados Unidos
Fil: Hawkins, Caleb W. D.. University of Georgia; Estados Unidos
Fil: Tarleton, Rick L.. University of Georgia; Estados Unidos
Materia
TRYPANOSOMA CRUZI
CHAGAS
CLEARING
IMMUNOSTAINING
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/212957
Acceder
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network_acronym_str CONICETDig
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network_name_str CONICET Digital (CONICET)
spelling Quantitative 3D Imaging of Trypanosoma cruzi-Infected Cells, Dormant Amastigotes, and T Cells in Intact Clarified OrgansSánchez Valdéz, Fernando JavierPadilla, Angel MarceloBustamante, Juan ManuelHawkins, Caleb W. D.Tarleton, Rick L.TRYPANOSOMA CRUZICHAGASCLEARINGIMMUNOSTAININGhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Reliable detection of Trypanosoma cruzi (T. cruzi) in vivo infections have long been needed to understand the complex biology of Chagas disease and to accurately evaluate the outcome of treatment regimens. Here, an integrated pipeline for automated quantification of T. cruzi-infected cells in 3D-reconstructed, cleared organs was developed. Light-sheet fluorescent microscopy allows us to accurately visualize and quantify not only actively proliferating but also dormant T. cruzi parasites and immune effector cells in whole-organs or tissues. Also, CUBIC-HistoVision pipeline to obtain uniform labeling of cleared organs with antibodies and nuclear stains was successfully adopted. Tissue clearing coupled to 3D immunostaining provides an unbiased approach to comprehensively evaluate drug treatment protocols, improve the understanding of the cellular organization of T. cruzi-infected tissues and is expected to advance discoveries related to anti-T. cruzi immune responses, tissue damage and repair in Chagas disease.Fil: Sánchez Valdéz, Fernando Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; ArgentinaFil: Padilla, Angel Marcelo. University of Georgia; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Bustamante, Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. University of Georgia; Estados UnidosFil: Hawkins, Caleb W. D.. University of Georgia; Estados UnidosFil: Tarleton, Rick L.. University of Georgia; Estados UnidosJournal of Visualized Experiments2021-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/212957Sánchez Valdéz, Fernando Javier; Padilla, Angel Marcelo; Bustamante, Juan Manuel; Hawkins, Caleb W. D.; Tarleton, Rick L.; Quantitative 3D Imaging of Trypanosoma cruzi-Infected Cells, Dormant Amastigotes, and T Cells in Intact Clarified Organs; Journal of Visualized Experiments; Journal of Visualized Experiments; 184; 7-2021; 1-181940-087XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.jove.com/es/v/63919/quantitative-3d-imaging-trypanosoma-cruzi-infected-cells-dormantinfo:eu-repo/semantics/altIdentifier/doi/10.3791/63919-vinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:04:33Zoai:ri.conicet.gov.ar:11336/212957instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:04:33.587CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Quantitative 3D Imaging of Trypanosoma cruzi-Infected Cells, Dormant Amastigotes, and T Cells in Intact Clarified Organs
title Quantitative 3D Imaging of Trypanosoma cruzi-Infected Cells, Dormant Amastigotes, and T Cells in Intact Clarified Organs
spellingShingle Quantitative 3D Imaging of Trypanosoma cruzi-Infected Cells, Dormant Amastigotes, and T Cells in Intact Clarified Organs
Sánchez Valdéz, Fernando Javier
TRYPANOSOMA CRUZI
CHAGAS
CLEARING
IMMUNOSTAINING
title_short Quantitative 3D Imaging of Trypanosoma cruzi-Infected Cells, Dormant Amastigotes, and T Cells in Intact Clarified Organs
title_full Quantitative 3D Imaging of Trypanosoma cruzi-Infected Cells, Dormant Amastigotes, and T Cells in Intact Clarified Organs
title_fullStr Quantitative 3D Imaging of Trypanosoma cruzi-Infected Cells, Dormant Amastigotes, and T Cells in Intact Clarified Organs
title_full_unstemmed Quantitative 3D Imaging of Trypanosoma cruzi-Infected Cells, Dormant Amastigotes, and T Cells in Intact Clarified Organs
title_sort Quantitative 3D Imaging of Trypanosoma cruzi-Infected Cells, Dormant Amastigotes, and T Cells in Intact Clarified Organs
dc.creator.none.fl_str_mv Sánchez Valdéz, Fernando Javier
Padilla, Angel Marcelo
Bustamante, Juan Manuel
Hawkins, Caleb W. D.
Tarleton, Rick L.
author Sánchez Valdéz, Fernando Javier
author_facet Sánchez Valdéz, Fernando Javier
Padilla, Angel Marcelo
Bustamante, Juan Manuel
Hawkins, Caleb W. D.
Tarleton, Rick L.
author_role author
author2 Padilla, Angel Marcelo
Bustamante, Juan Manuel
Hawkins, Caleb W. D.
Tarleton, Rick L.
author2_role author
author
author
author
dc.subject.none.fl_str_mv TRYPANOSOMA CRUZI
CHAGAS
CLEARING
IMMUNOSTAINING
topic TRYPANOSOMA CRUZI
CHAGAS
CLEARING
IMMUNOSTAINING
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Reliable detection of Trypanosoma cruzi (T. cruzi) in vivo infections have long been needed to understand the complex biology of Chagas disease and to accurately evaluate the outcome of treatment regimens. Here, an integrated pipeline for automated quantification of T. cruzi-infected cells in 3D-reconstructed, cleared organs was developed. Light-sheet fluorescent microscopy allows us to accurately visualize and quantify not only actively proliferating but also dormant T. cruzi parasites and immune effector cells in whole-organs or tissues. Also, CUBIC-HistoVision pipeline to obtain uniform labeling of cleared organs with antibodies and nuclear stains was successfully adopted. Tissue clearing coupled to 3D immunostaining provides an unbiased approach to comprehensively evaluate drug treatment protocols, improve the understanding of the cellular organization of T. cruzi-infected tissues and is expected to advance discoveries related to anti-T. cruzi immune responses, tissue damage and repair in Chagas disease.
Fil: Sánchez Valdéz, Fernando Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina
Fil: Padilla, Angel Marcelo. University of Georgia; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Bustamante, Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. University of Georgia; Estados Unidos
Fil: Hawkins, Caleb W. D.. University of Georgia; Estados Unidos
Fil: Tarleton, Rick L.. University of Georgia; Estados Unidos
description Reliable detection of Trypanosoma cruzi (T. cruzi) in vivo infections have long been needed to understand the complex biology of Chagas disease and to accurately evaluate the outcome of treatment regimens. Here, an integrated pipeline for automated quantification of T. cruzi-infected cells in 3D-reconstructed, cleared organs was developed. Light-sheet fluorescent microscopy allows us to accurately visualize and quantify not only actively proliferating but also dormant T. cruzi parasites and immune effector cells in whole-organs or tissues. Also, CUBIC-HistoVision pipeline to obtain uniform labeling of cleared organs with antibodies and nuclear stains was successfully adopted. Tissue clearing coupled to 3D immunostaining provides an unbiased approach to comprehensively evaluate drug treatment protocols, improve the understanding of the cellular organization of T. cruzi-infected tissues and is expected to advance discoveries related to anti-T. cruzi immune responses, tissue damage and repair in Chagas disease.
publishDate 2021
dc.date.none.fl_str_mv 2021-07
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/212957
Sánchez Valdéz, Fernando Javier; Padilla, Angel Marcelo; Bustamante, Juan Manuel; Hawkins, Caleb W. D.; Tarleton, Rick L.; Quantitative 3D Imaging of Trypanosoma cruzi-Infected Cells, Dormant Amastigotes, and T Cells in Intact Clarified Organs; Journal of Visualized Experiments; Journal of Visualized Experiments; 184; 7-2021; 1-18
1940-087X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/212957
identifier_str_mv Sánchez Valdéz, Fernando Javier; Padilla, Angel Marcelo; Bustamante, Juan Manuel; Hawkins, Caleb W. D.; Tarleton, Rick L.; Quantitative 3D Imaging of Trypanosoma cruzi-Infected Cells, Dormant Amastigotes, and T Cells in Intact Clarified Organs; Journal of Visualized Experiments; Journal of Visualized Experiments; 184; 7-2021; 1-18
1940-087X
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.jove.com/es/v/63919/quantitative-3d-imaging-trypanosoma-cruzi-infected-cells-dormant
info:eu-repo/semantics/altIdentifier/doi/10.3791/63919-v
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Journal of Visualized Experiments
publisher.none.fl_str_mv Journal of Visualized Experiments
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1844613872733913088
score 13.070432

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