Development of an electrochemical microsensor for monitoring the respiration activity of single cells and animals by scanning electrochemical microscopy

Detalhes bibliográficos
Ano de defesa: 2019
Autor(a) principal: Santos, Carla Santana
Orientador(a): Não Informado pela instituição
Banca de defesa: Não Informado pela instituição
Tipo de documento: Tese
Tipo de acesso: Acesso aberto
Idioma: eng
Instituição de defesa: Biblioteca Digitais de Teses e Dissertações da USP
Programa de Pós-Graduação: Não Informado pela instituição
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Palavras-chave em Português:
OCR
Link de acesso: https://www.teses.usp.br/teses/disponiveis/46/46136/tde-09032020-145525/
Resumo: This thesis shows results on the development of an O2 micro/nanosensor for scanning electrochemical microscopy (SECM) application with the aim of monitoring changes in respiration activity over a single cell. The surface of platinum microelectrodes was modified by using a platinisation protocol in order to facilitate the eletron-transfer process regarding the O2 reduction reaction, hence the sensitivity of the device was greatly enhanced. The stability of the microsensor and the nature of oxygen reduction reaction (ORR) products were also evaluated by SECM. The developed microsensor was employed in redox competition SECM operating mode to monitor local changes in the O2 concentration of a single-cell (HS578T lineage) present in phosphate buffer medium during the addition of specific compounds to modulate the mitochondria activity. A similar protocol was used for examining the metabolism of a nematode (Caenorhabditis elegans) by scanning the O2 microsensor over a single-animal, with particular attention being devoted to the reproductive system. Hence, SECM has been proven to be a powerful and reliable technique to investigate biochemical events related to cellular activity by means of oxygen consumption rate (OCR) with high spatial resolution. The last section of this thesis presents a novel scanning probe microscopy developed in the Prof. Schuhmanns group (Germany) employing bipolar electrochemistry concepts. The scanning bipolar electrochemical microscopy (SBECM) allows the independent acquisition of multi-microelectrodes responses localized close to a surface. In contrast of scanning a single probe as the conventional SECM, higher temporal resolution is obtained with SBECM. The results showed in this thesis confirmed the ability of the electrochemical probe techniques to acquire local O2 concentration information and to provide deeper insight and comprehensive aspects in the biological area.
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spelling Development of an electrochemical microsensor for monitoring the respiration activity of single cells and animals by scanning electrochemical microscopyDesenvolvimento de microssensor eletroquímico para o monitoramento da atividade respiratória de células e animais por microscopia eletroquímica de varreduraBipolar electrochemistryEletroquímica bipolaMicroelectrodeMicroeletrodoMicroscopia EletroquímicaOCROCROxygen sensorScanning Electrochemical MicroscopySECMSECMSensor de oxigênioThis thesis shows results on the development of an O2 micro/nanosensor for scanning electrochemical microscopy (SECM) application with the aim of monitoring changes in respiration activity over a single cell. The surface of platinum microelectrodes was modified by using a platinisation protocol in order to facilitate the eletron-transfer process regarding the O2 reduction reaction, hence the sensitivity of the device was greatly enhanced. The stability of the microsensor and the nature of oxygen reduction reaction (ORR) products were also evaluated by SECM. The developed microsensor was employed in redox competition SECM operating mode to monitor local changes in the O2 concentration of a single-cell (HS578T lineage) present in phosphate buffer medium during the addition of specific compounds to modulate the mitochondria activity. A similar protocol was used for examining the metabolism of a nematode (Caenorhabditis elegans) by scanning the O2 microsensor over a single-animal, with particular attention being devoted to the reproductive system. Hence, SECM has been proven to be a powerful and reliable technique to investigate biochemical events related to cellular activity by means of oxygen consumption rate (OCR) with high spatial resolution. The last section of this thesis presents a novel scanning probe microscopy developed in the Prof. Schuhmanns group (Germany) employing bipolar electrochemistry concepts. The scanning bipolar electrochemical microscopy (SBECM) allows the independent acquisition of multi-microelectrodes responses localized close to a surface. In contrast of scanning a single probe as the conventional SECM, higher temporal resolution is obtained with SBECM. The results showed in this thesis confirmed the ability of the electrochemical probe techniques to acquire local O2 concentration information and to provide deeper insight and comprehensive aspects in the biological area.Nesta tese são apresentados resultados sobre o desenvolvimento de um micro/nanossensor de O2 para aplicação em microscopia eletroquímica de varredura (SECM) com o objetivo de monitorar alterações na atividade respiratória em uma única célula. A superfície dos microeletrodos de platina foi modificada por meio de um protocolo de platinização, visando à facilitação do processo de transferência de elétrons da reação de redução do O2. Como consequência, a sensibilidade do dispositivo no que diz respeito à detecção de O2 foi bastante aumentada. A estabilidade do microssensor e a natureza dos produtos ORR também foram avaliadas por SECM. O microssensor desenvolvido foi empregado no modo de competição redox da SECM para monitorar alterações locais na concentração de O2 em células (linhagem HS578T) em tampão fosfato durante a adição de substâncias específicas para modular a atividade das mitocôndrias. Um protocolo semelhante foi usado para examinar o metabolismo de um nemátodo (Caenorhabditis elegans) pela movimentação do microssensor de O2 sobre um único animal, com atenção especial sendo dedicada ao sistema reprodutivo. Portanto, a SECM demonstrou ser uma técnica poderosa e confiável para investigar eventos bioquímicos relacionados à atividade celular com alta resolução espacial. A última seção desta tese apresenta uma nova modalidade de microscopia de varredura desenvolvida no grupo do Prof. Schuhmann (Alemanha), baseada em conceitos de eletroquímica bipolar. A microscopia eletroquímica bipolar de varredura (SBECM) permite a aquisição independente de respostas de multi-microeletrodos localizados próximos a uma superfície. Ao contrário do deslocamento de uma única sonda como na SECM convencional, no caso da SBECM dados são obtidos com maior resolução temporal. Os resultados apresentados nesta tese confirmaram a versatilidade das técnicas de sondas eletroquímicas para a obtenção de informações localizadas sobre a concentração de oxigênio e como ferramentas para a expansão de conhecimentos mais aprofundados e abrangentes na área biológica.Biblioteca Digitais de Teses e Dissertações da USPBertotti, MauroSantos, Carla Santana2019-12-02info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttps://www.teses.usp.br/teses/disponiveis/46/46136/tde-09032020-145525/reponame:Biblioteca Digital de Teses e Dissertações da USPinstname:Universidade de São Paulo (USP)instacron:USPLiberar o conteúdo para acesso público.info:eu-repo/semantics/openAccesseng2020-08-08T00:04:02Zoai:teses.usp.br:tde-09032020-145525Biblioteca Digital de Teses e Dissertaçõeshttp://www.teses.usp.br/PUBhttp://www.teses.usp.br/cgi-bin/mtd2br.plvirginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.bropendoar:27212020-08-08T00:04:02Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)false
dc.title.none.fl_str_mv Development of an electrochemical microsensor for monitoring the respiration activity of single cells and animals by scanning electrochemical microscopy
Desenvolvimento de microssensor eletroquímico para o monitoramento da atividade respiratória de células e animais por microscopia eletroquímica de varredura
title Development of an electrochemical microsensor for monitoring the respiration activity of single cells and animals by scanning electrochemical microscopy
spellingShingle Development of an electrochemical microsensor for monitoring the respiration activity of single cells and animals by scanning electrochemical microscopy
Santos, Carla Santana
Bipolar electrochemistry
Eletroquímica bipola
Microelectrode
Microeletrodo
Microscopia Eletroquímica
OCR
OCR
Oxygen sensor
Scanning Electrochemical Microscopy
SECM
SECM
Sensor de oxigênio
title_short Development of an electrochemical microsensor for monitoring the respiration activity of single cells and animals by scanning electrochemical microscopy
title_full Development of an electrochemical microsensor for monitoring the respiration activity of single cells and animals by scanning electrochemical microscopy
title_fullStr Development of an electrochemical microsensor for monitoring the respiration activity of single cells and animals by scanning electrochemical microscopy
title_full_unstemmed Development of an electrochemical microsensor for monitoring the respiration activity of single cells and animals by scanning electrochemical microscopy
title_sort Development of an electrochemical microsensor for monitoring the respiration activity of single cells and animals by scanning electrochemical microscopy
author Santos, Carla Santana
author_facet Santos, Carla Santana
author_role author
dc.contributor.none.fl_str_mv Bertotti, Mauro
dc.contributor.author.fl_str_mv Santos, Carla Santana
dc.subject.por.fl_str_mv Bipolar electrochemistry
Eletroquímica bipola
Microelectrode
Microeletrodo
Microscopia Eletroquímica
OCR
OCR
Oxygen sensor
Scanning Electrochemical Microscopy
SECM
SECM
Sensor de oxigênio
topic Bipolar electrochemistry
Eletroquímica bipola
Microelectrode
Microeletrodo
Microscopia Eletroquímica
OCR
OCR
Oxygen sensor
Scanning Electrochemical Microscopy
SECM
SECM
Sensor de oxigênio
description This thesis shows results on the development of an O2 micro/nanosensor for scanning electrochemical microscopy (SECM) application with the aim of monitoring changes in respiration activity over a single cell. The surface of platinum microelectrodes was modified by using a platinisation protocol in order to facilitate the eletron-transfer process regarding the O2 reduction reaction, hence the sensitivity of the device was greatly enhanced. The stability of the microsensor and the nature of oxygen reduction reaction (ORR) products were also evaluated by SECM. The developed microsensor was employed in redox competition SECM operating mode to monitor local changes in the O2 concentration of a single-cell (HS578T lineage) present in phosphate buffer medium during the addition of specific compounds to modulate the mitochondria activity. A similar protocol was used for examining the metabolism of a nematode (Caenorhabditis elegans) by scanning the O2 microsensor over a single-animal, with particular attention being devoted to the reproductive system. Hence, SECM has been proven to be a powerful and reliable technique to investigate biochemical events related to cellular activity by means of oxygen consumption rate (OCR) with high spatial resolution. The last section of this thesis presents a novel scanning probe microscopy developed in the Prof. Schuhmanns group (Germany) employing bipolar electrochemistry concepts. The scanning bipolar electrochemical microscopy (SBECM) allows the independent acquisition of multi-microelectrodes responses localized close to a surface. In contrast of scanning a single probe as the conventional SECM, higher temporal resolution is obtained with SBECM. The results showed in this thesis confirmed the ability of the electrochemical probe techniques to acquire local O2 concentration information and to provide deeper insight and comprehensive aspects in the biological area.
publishDate 2019
dc.date.none.fl_str_mv 2019-12-02
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://www.teses.usp.br/teses/disponiveis/46/46136/tde-09032020-145525/
url https://www.teses.usp.br/teses/disponiveis/46/46136/tde-09032020-145525/
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv
dc.rights.driver.fl_str_mv Liberar o conteúdo para acesso público.
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Liberar o conteúdo para acesso público.
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.coverage.none.fl_str_mv
dc.publisher.none.fl_str_mv Biblioteca Digitais de Teses e Dissertações da USP
publisher.none.fl_str_mv Biblioteca Digitais de Teses e Dissertações da USP
dc.source.none.fl_str_mv
reponame:Biblioteca Digital de Teses e Dissertações da USP
instname:Universidade de São Paulo (USP)
instacron:USP
instname_str Universidade de São Paulo (USP)
instacron_str USP
institution USP
reponame_str Biblioteca Digital de Teses e Dissertações da USP
collection Biblioteca Digital de Teses e Dissertações da USP
repository.name.fl_str_mv Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)
repository.mail.fl_str_mv virginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.br
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