Development of an electrochemical microsensor for monitoring the respiration activity of single cells and animals by scanning electrochemical microscopy
Ano de defesa: | 2019 |
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Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | |
Tipo de documento: | Tese |
Tipo de acesso: | Acesso aberto |
Idioma: | eng |
Instituição de defesa: |
Biblioteca Digitais de Teses e Dissertações da USP
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Programa de Pós-Graduação: |
Não Informado pela instituição
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Departamento: |
Não Informado pela instituição
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País: |
Não Informado pela instituição
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Palavras-chave em Português: | |
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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Biblioteca Digital de Teses e Dissertações da USP |
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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 |
_version_ |
1815257997850443776 |