Development of lens-free holographic microscopes using multiheight and multispectral phase recovery methods
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Tipo de documento: | Tese |
Idioma: | eng |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da USP |
Texto Completo: | https://www.teses.usp.br/teses/disponiveis/76/76134/tde-29052023-143103/ |
Resumo: | Since its invention in century XVI, microscopy is one of the most important optics techniques. Its use has revolutionized many knowledge areas and these applications have driven improvements in the technique itself. The lens-free microscope has emerged as an alternative to traditional optical microscopes. This recent technique has the advantage to decouple the imaging resolution from the field of view and also to provide very compact and low-cost systems. Lens-free microscopes based on hologram acquisition produce amplitude and phase images of the same observed scene, using a digital imaging process to perform phase recovery. This study aims to develop and test portable and automated lens-free holographic microscopes, based on open-source software, using two methods for imaging acquisition and processing: multiheight and multispectral. The first applied method is the multiheight, which records a sequence of holograms with different sensor-sample distances and uses them as input data to recover phase values of the light that better describes the recorded scene. Multispectral method was also used to record holograms using light with emission at different spectral ranges and composes amplitude and phase images from this imaging information. Homemade open-source algorithms were developed in Python language to control the component device and to perform the digital imaging processing. Both lens-free microscopes were tested using standard test targets to define the spatial resolution of each one. Besides, it has been applied to imaging different types of biological samples to analyze their amplitude and phase images in different conditions. Thus, these systems were used to image different cells and microorganisms. More robust analyses were performed to provide yeast and Giardia cyst counting. Therefore, two portable, low-cost, automated, and based on open-source software lens-free holographic microscopes were developed and tested in this study, using multiheight and multispectral methods to perform amplitude and phase images with almost 30mm2 and about 5μm of resolution. Both systems proved to be efficient for observing cells and microorganisms without the need for labels. |
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Development of lens-free holographic microscopes using multiheight and multispectral phase recovery methodsDesenvolvimento de microscópios holográficos sem lentes usando métodos de recuperação de fase multialtura e multiespectralCódigo abertoLensless optical microscopyMicroscópio óptico sem lentesMultialturasMultiespectralMultiheightMultispectralOpensourcePhase recoveryRecuperação de faseSince its invention in century XVI, microscopy is one of the most important optics techniques. Its use has revolutionized many knowledge areas and these applications have driven improvements in the technique itself. The lens-free microscope has emerged as an alternative to traditional optical microscopes. This recent technique has the advantage to decouple the imaging resolution from the field of view and also to provide very compact and low-cost systems. Lens-free microscopes based on hologram acquisition produce amplitude and phase images of the same observed scene, using a digital imaging process to perform phase recovery. This study aims to develop and test portable and automated lens-free holographic microscopes, based on open-source software, using two methods for imaging acquisition and processing: multiheight and multispectral. The first applied method is the multiheight, which records a sequence of holograms with different sensor-sample distances and uses them as input data to recover phase values of the light that better describes the recorded scene. Multispectral method was also used to record holograms using light with emission at different spectral ranges and composes amplitude and phase images from this imaging information. Homemade open-source algorithms were developed in Python language to control the component device and to perform the digital imaging processing. Both lens-free microscopes were tested using standard test targets to define the spatial resolution of each one. Besides, it has been applied to imaging different types of biological samples to analyze their amplitude and phase images in different conditions. Thus, these systems were used to image different cells and microorganisms. More robust analyses were performed to provide yeast and Giardia cyst counting. Therefore, two portable, low-cost, automated, and based on open-source software lens-free holographic microscopes were developed and tested in this study, using multiheight and multispectral methods to perform amplitude and phase images with almost 30mm2 and about 5μm of resolution. Both systems proved to be efficient for observing cells and microorganisms without the need for labels.Desde a sua invenção no século XVI, a microscopia é uma das técnicas ópticas mais importantes. Seu uso revolucionou muitas áreas do conhecimento e essas aplicações têm impulsionado melhorias na própria técnica. O microscópio sem lentes surgiu como uma alternativa aos microscópios ópticos tradicionais. Esta técnica recente tem a vantagem de desacoplar a resolução da imagem do campo de visão, além de fornecer sistemas muito compactos e de baixo custo. Os microscópios sem lente baseados na aquisição de holograma produzem imagens de amplitude e fase da mesma cena observada, usando um processamento digital de imagem para realizar a recuperação de fase. Este estudo tem como objetivo desenvolver e testar microscópios holográficos sem lentes portáteis e automatizados, baseados em algoritmos computacionais de códigos abertos, aplicando dois métodos para aquisição e processamento das imagens: multialturas e multiespectral. O primeiro método aplicado é o multialturas, que registra uma sequência de hologramas com diferentes distâncias entre o sensor e a amostra e os utiliza como dados de entrada para recuperar os valores de fase da luz que melhor descrevem a cena gravada. O método multiespectral também foi usado para registrar hologramas, usando luz com emissão em diferentes faixas espectrais para compor imagens de amplitude e fase a partir dessas informações em forma de imagem. Algoritmos de código aberto foram desenvolvidos em linguagem Python para controlar os dispositivos que compõem o sistema e realizar os processamentos digitais das imagens. Ambos os microscópios sem lentes foram testados usando alvo de teste padrão para definir a resolução espacial de cada um. Além disso, eles foram aplicados para observar diferentes tipos de amostras biológicas, de modo a analisar suas imagens de amplitude e fase em condições distintas. Assim, esses microscópios foram usados para observar diferentes tipos de células e microorganismos. Análises mais robustas foram realizadas para fornecer mais informações a respeito das amostras. Portanto, dois microscópios holográficos sem lentes portáteis, de baixo custo, automatizados e baseados em algoritmos computacionais de código aberto foram desenvolvidos e testados neste estudo, usando métodos multialturas e multiespectrais para realizar imagens de amplitude e fase com quase 30mm2 e cerca de 5μm de resolução. Ambos sistemas se mostraram eficientes para observar células e microorganismos sem a necessidade de marcação.Biblioteca Digitais de Teses e Dissertações da USPPratavieira, SebastiãoD\'Almeida, Camila de Paula2023-03-28info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttps://www.teses.usp.br/teses/disponiveis/76/76134/tde-29052023-143103/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/openAccesseng2024-08-23T15:31:02Zoai:teses.usp.br:tde-29052023-143103Biblioteca 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:27212024-08-23T15:31:02Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)false |
dc.title.none.fl_str_mv |
Development of lens-free holographic microscopes using multiheight and multispectral phase recovery methods Desenvolvimento de microscópios holográficos sem lentes usando métodos de recuperação de fase multialtura e multiespectral |
title |
Development of lens-free holographic microscopes using multiheight and multispectral phase recovery methods |
spellingShingle |
Development of lens-free holographic microscopes using multiheight and multispectral phase recovery methods D\'Almeida, Camila de Paula Código aberto Lensless optical microscopy Microscópio óptico sem lentes Multialturas Multiespectral Multiheight Multispectral Opensource Phase recovery Recuperação de fase |
title_short |
Development of lens-free holographic microscopes using multiheight and multispectral phase recovery methods |
title_full |
Development of lens-free holographic microscopes using multiheight and multispectral phase recovery methods |
title_fullStr |
Development of lens-free holographic microscopes using multiheight and multispectral phase recovery methods |
title_full_unstemmed |
Development of lens-free holographic microscopes using multiheight and multispectral phase recovery methods |
title_sort |
Development of lens-free holographic microscopes using multiheight and multispectral phase recovery methods |
author |
D\'Almeida, Camila de Paula |
author_facet |
D\'Almeida, Camila de Paula |
author_role |
author |
dc.contributor.none.fl_str_mv |
Pratavieira, Sebastião |
dc.contributor.author.fl_str_mv |
D\'Almeida, Camila de Paula |
dc.subject.por.fl_str_mv |
Código aberto Lensless optical microscopy Microscópio óptico sem lentes Multialturas Multiespectral Multiheight Multispectral Opensource Phase recovery Recuperação de fase |
topic |
Código aberto Lensless optical microscopy Microscópio óptico sem lentes Multialturas Multiespectral Multiheight Multispectral Opensource Phase recovery Recuperação de fase |
description |
Since its invention in century XVI, microscopy is one of the most important optics techniques. Its use has revolutionized many knowledge areas and these applications have driven improvements in the technique itself. The lens-free microscope has emerged as an alternative to traditional optical microscopes. This recent technique has the advantage to decouple the imaging resolution from the field of view and also to provide very compact and low-cost systems. Lens-free microscopes based on hologram acquisition produce amplitude and phase images of the same observed scene, using a digital imaging process to perform phase recovery. This study aims to develop and test portable and automated lens-free holographic microscopes, based on open-source software, using two methods for imaging acquisition and processing: multiheight and multispectral. The first applied method is the multiheight, which records a sequence of holograms with different sensor-sample distances and uses them as input data to recover phase values of the light that better describes the recorded scene. Multispectral method was also used to record holograms using light with emission at different spectral ranges and composes amplitude and phase images from this imaging information. Homemade open-source algorithms were developed in Python language to control the component device and to perform the digital imaging processing. Both lens-free microscopes were tested using standard test targets to define the spatial resolution of each one. Besides, it has been applied to imaging different types of biological samples to analyze their amplitude and phase images in different conditions. Thus, these systems were used to image different cells and microorganisms. More robust analyses were performed to provide yeast and Giardia cyst counting. Therefore, two portable, low-cost, automated, and based on open-source software lens-free holographic microscopes were developed and tested in this study, using multiheight and multispectral methods to perform amplitude and phase images with almost 30mm2 and about 5μm of resolution. Both systems proved to be efficient for observing cells and microorganisms without the need for labels. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023-03-28 |
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/76/76134/tde-29052023-143103/ |
url |
https://www.teses.usp.br/teses/disponiveis/76/76134/tde-29052023-143103/ |
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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1809091175130333184 |