On-chip biosensing platforms based on gold and silicon optical nano-resonators

Point-of-care (POC) devices are compact, mobile and fast detection platforms expected to advance early diagnosis, treatment monitoring and personalized healthcare, and revolutionize today’s healthcare system, especially in remote areas. The need for POC devices strongly drives the development of novel biosensor technology. Building a small, fast, simple, and sensitive platform for biomolecule detection is a challenge that relies on the integration of multiple fields of expertise and engineering. Optical nanoresonators have shown great promise as label-free biosensors because of direct light coupling and sub-wavelength sensing modes. Metallic nanoresonators with localized surface plasmon resonances (LSPR) are already well studied and were proven a solid alternative to the commercialized surface plasmon resonance (SPR) sensors. More recently, dielectric nanoresonators have also gained traction due to the reduced losses and the ability to manipulate both the electric and magnetic components of the incident light. In this thesis, we advance the field of biosensing and use optical nanoresonators as operative platforms relevant for disease diagnosis and treatment monitoring. By combining different optimized optical nanoresonators, both metallic and dielectric, with state-of-the-art microfluidics and surface chemistry, we have developed and tested several detection platforms. We first focused on developing a microfluidic lab-on-chip device for multiplexed biosensing utilizing the LSPR of gold nanoresonator arrays. By simultaneously tracking the extinction of 32 sensor arrays, we demonstrated multiplexed quantitative detection of four breast cancer markers in human serum. We showed that with well-optimized immunoassays, a low limit of detection (LOD) can be reached, paving the way towards clinically-relevant POC devices. Additionally, we implemented silicon nanoresonators supporting Mie resonances into functional and clinically-relevant applications. By integrating several arrays of Si nanoresonators with state-of-the-art microfluidics, we demonstrated their ability to detect cancer markers in human serum with high sensitivity and high specificity. Furthermore, we showed that the fabrication of Si nanoresonator array using low cost and scalable projection lithography leads to sufficiently low limits of detection, while enabling cheaper and faster sensor production for future POC applications. We also investigated the respective role of electric and magnetic dipole resonances and showed that they are associated with two different transduction mechanisms: resonance redshift and extinction decrease. Our work advances the development of future point-of-care sensing platforms for fast and low cost health monitoring at the molecular scale.La instrumentación Point-of-care (POC) es compacta, móvil y permite una detección rápida, razón por la que se prevé que sean de gran ayuda en áreas como el diagnostico precoz, la monitorización de tratamientos y la medicina personalizada, revolucionando los modelos sanitarios, especialmente en las zonas de difícil acceso y con menos recursos. La necesidad de este tipo de dispositivos impulsa el desarrollo de novedosas tecnologías en el campo de los bio-sensores. Diseñar equipos para la detección de bio-moléculas que sean rápidos, pequeños y sencillos es un reto que requiere la integración de múltiples campos de la ciencia y la ingeniería. Los nano-resonadores ópticos muestran un gran potencial como bio-sensores sin necesidad de marcaje, gracias a su capacidad de acoplase directamente con la luz en modos menores que la longitud de onda. Los nano-resonadores metálicos basados en resonancias plasmónicas superficiales localizadas (LSPR) han sido estudiados y han demostrado ser una firme alternativa a los ya comerciales basados en resonancias plasmónicas superficiales (SPR). Los nano-resonadores dieléctricos han sido recientemente objeto de atención debido a sus bajas perdidas y la capacidad de manipular los componentes eléctricos y magnéticos de la luz. En esta tesis presentamos avances en el campo de la bio-detección y en el uso de los nano-resonadores ópticos como potenciales herramientas para la detección de enfermedades y monitorización de los tratamientos. Hemos desarrollado y evaluado distintas plataformas de detección combinando los nano-resonadores ópticos, tanto metálicos como dieléctricos, con las más avanzadas técnicas de microfluídica y química de superficies. En primer lugar, nos centramos en el desarrollo de un dispositivo microfluídico basado en sensores LSPR de oro que permite multiplexar 32 canales. Los 32 sensores se monitorizan en tiempo real para demostrar la cuantificación de 4 marcadores de cáncer de mama en suero sanguíneo humano. Demostramos que mediante la optimización de los ensayos se pueden alcanzar bajos límites de detección (LOD), lo que allana el camino hacia dispositivos POC de uso clínico. Por otro lado, hemos utilizado los nano-resonadores de silicio integrados con la microfluídica para también detectar marcadores de cáncer en suero. Estos sensores, cuyo principio de funcionamiento se basa en resonancias de MIE, han demostrado ser una alternativa razonable a los sensores de oro. Además, demostramos que un proceso de fabricación de nano-resonadores de silicio rápido, escalable y de bajo coste da lugar a límites de detección suficientes para la producción de futuras POC. También realizamos un minucioso estudio del rol de las resonancias eléctricas y magnéticas en dichos sensores y su relación con el desplazamiento y el cambio magnitud de la resonancia del sensor global. Nuestro trabajo es un avance en el desarrollo de futuros instrumentos POC rápidos y baratos en el ámbito de la salud a escala molecular.Postprint (published version

On-a-chip biosensing based on all-dielectric nanoresonators

Nanophotonics has become a key enabling technology in biomedicine with great promises in early diagnosis and less invasive therapies. In this context, the unique capability of plasmonic noble metal nanoparticles to concentrate light on the nanometer scale has widely contributed to biosensing and enhanced spectroscopy. Recently, high-refractive index dielectric nanostructures featuring low loss resonances have been proposed as a promising alternative to nanoplasmonics, potentially offering better sensing performances along with full compatibility with the microelectronics industry. In this letter we report the first demonstration of biosensing with silicon nanoresonators integrated in state-of-the-art microfluidics. Our lab-on-a-chip platform enables detecting Prostate Specific Antigen (PSA) cancer marker in human serum with a sensitivity that meets clinical needs. These performances are directly compared with its plasmonic counterpart based on gold nanorods. Our work opens new opportunities in the development of future point-of-care devices towards a more personalized healthcare

On-chip biosensing platforms based on gold and silicon optical nano-resonators

Point-of-care (POC) devices are compact, mobile and fast detection platforms expected to advance early diagnosis, treatment monitoring and personalized healthcare, and revolutionize today’s healthcare system, especially in remote areas. The need for POC devices strongly drives the development of novel biosensor technology. Building a small, fast, simple, and sensitive platform for biomolecule detection is a challenge that relies on the integration of multiple fields of expertise and engineering. Optical nanoresonators have shown great promise as label-free biosensors because of direct light coupling and sub-wavelength sensing modes. Metallic nanoresonators with localized surface plasmon resonances (LSPR) are already well studied and were proven a solid alternative to the commercialized surface plasmon resonance (SPR) sensors. More recently, dielectric nanoresonators have also gained traction due to the reduced losses and the ability to manipulate both the electric and magnetic components of the incident light. In this thesis, we advance the field of biosensing and use optical nanoresonators as operative platforms relevant for disease diagnosis and treatment monitoring. By combining different optimized optical nanoresonators, both metallic and dielectric, with state-of-the-art microfluidics and surface chemistry, we have developed and tested several detection platforms. We first focused on developing a microfluidic lab-on-chip device for multiplexed biosensing utilizing the LSPR of gold nanoresonator arrays. By simultaneously tracking the extinction of 32 sensor arrays, we demonstrated multiplexed quantitative detection of four breast cancer markers in human serum. We showed that with well-optimized immunoassays, a low limit of detection (LOD) can be reached, paving the way towards clinically-relevant POC devices. Additionally, we implemented silicon nanoresonators supporting Mie resonances into functional and clinically-relevant applications. By integrating several arrays of Si nanoresonators with state-of-the-art microfluidics, we demonstrated their ability to detect cancer markers in human serum with high sensitivity and high specificity. Furthermore, we showed that the fabrication of Si nanoresonator array using low cost and scalable projection lithography leads to sufficiently low limits of detection, while enabling cheaper and faster sensor production for future POC applications. We also investigated the respective role of electric and magnetic dipole resonances and showed that they are associated with two different transduction mechanisms: resonance redshift and extinction decrease. Our work advances the development of future point-of-care sensing platforms for fast and low cost health monitoring at the molecular scale.La instrumentación Point-of-care (POC) es compacta, móvil y permite una detección rápida, razón por la que se prevé que sean de gran ayuda en áreas como el diagnostico precoz, la monitorización de tratamientos y la medicina personalizada, revolucionando los modelos sanitarios, especialmente en las zonas de difícil acceso y con menos recursos. La necesidad de este tipo de dispositivos impulsa el desarrollo de novedosas tecnologías en el campo de los bio-sensores. Diseñar equipos para la detección de bio-moléculas que sean rápidos, pequeños y sencillos es un reto que requiere la integración de múltiples campos de la ciencia y la ingeniería. Los nano-resonadores ópticos muestran un gran potencial como bio-sensores sin necesidad de marcaje, gracias a su capacidad de acoplase directamente con la luz en modos menores que la longitud de onda. Los nano-resonadores metálicos basados en resonancias plasmónicas superficiales localizadas (LSPR) han sido estudiados y han demostrado ser una firme alternativa a los ya comerciales basados en resonancias plasmónicas superficiales (SPR). Los nano-resonadores dieléctricos han sido recientemente objeto de atención debido a sus bajas perdidas y la capacidad de manipular los componentes eléctricos y magnéticos de la luz. En esta tesis presentamos avances en el campo de la bio-detección y en el uso de los nano-resonadores ópticos como potenciales herramientas para la detección de enfermedades y monitorización de los tratamientos. Hemos desarrollado y evaluado distintas plataformas de detección combinando los nano-resonadores ópticos, tanto metálicos como dieléctricos, con las más avanzadas técnicas de microfluídica y química de superficies. En primer lugar, nos centramos en el desarrollo de un dispositivo microfluídico basado en sensores LSPR de oro que permite multiplexar 32 canales. Los 32 sensores se monitorizan en tiempo real para demostrar la cuantificación de 4 marcadores de cáncer de mama en suero sanguíneo humano. Demostramos que mediante la optimización de los ensayos se pueden alcanzar bajos límites de detección (LOD), lo que allana el camino hacia dispositivos POC de uso clínico. Por otro lado, hemos utilizado los nano-resonadores de silicio integrados con la microfluídica para también detectar marcadores de cáncer en suero. Estos sensores, cuyo principio de funcionamiento se basa en resonancias de MIE, han demostrado ser una alternativa razonable a los sensores de oro. Además, demostramos que un proceso de fabricación de nano-resonadores de silicio rápido, escalable y de bajo coste da lugar a límites de detección suficientes para la producción de futuras POC. También realizamos un minucioso estudio del rol de las resonancias eléctricas y magnéticas en dichos sensores y su relación con el desplazamiento y el cambio magnitud de la resonancia del sensor global. Nuestro trabajo es un avance en el desarrollo de futuros instrumentos POC rápidos y baratos en el ámbito de la salud a escala molecular

Unravelling the Role of Electric and Magnetic Dipoles in Biosensing with Si Nanoresonators

High refractive index dielectric nanoresonators are attracting much attention due to their ability to control both electric and magnetic components of light. Combining confined modes with reduced absorption losses, they have recently been proposed as an alternative to nanoplasmonic biosensors. In this context, we study the use of semi-random silicon nanocylinder arrays, fabricated with simple and scalable colloidal lithography for the efficient and reliable detection of biomolecules in biological samples. Interestingly, electric and magnetic dipole resonances are associated to two different transduction mechanisms: extinction decrease and resonance redshift, respectively. By contrasting both observables, we identify clear advantages in tracking changes in the extinction magnitude. Our data demonstrate that, despite its simplicity, the proposed platform is able to detect prostate specific antigen (PSA) in human serum with limits of detection meeting clinical needs.Peer ReviewedPostprint (author's final draft

In Situ LSPR Sensing of Secreted Insulin in Organ-on-Chip

Organ-on-a-chip (OOC) devices offer new approaches for metabolic disease modeling and drug discovery by providing biologically relevant models of tissues and organs in vitro with a high degree of control over experimental variables for high-content screening applications. Yet, to fully exploit the potential of these platforms, there is a need to interface them with integrated non-labeled sensing modules, capable of monitoring, in situ, their biochemical response to external stimuli, such as stress or drugs. In order to meet this need, we aim here to develop an integrated technology based on coupling a localized surface plasmon resonance (LSPR) sensing module to an OOC device to monitor the insulin in situ secretion in pancreatic islets, a key physiological event that is usually perturbed in metabolic diseases such as type 2 diabetes (T2D). As a proof of concept, we developed a biomimetic islet-on-a-chip (IOC) device composed of mouse pancreatic islets hosted in a cellulose-based scaffold as a novel approach. The IOC was interfaced with a state-of-the-art on-chip LSPR sensing platform to monitor the in situ insulin secretion. The developed platform offers a powerful tool to enable the in situ response study of microtissues to external stimuli for applications such as a drug-screening platform for human models, bypassing animal testing

On-chip biosensing platforms based on gold and silicon optical nano-resonators

Point-of-care (POC) devices are compact, mobile and fast detection platforms expected to advance early diagnosis, treatment monitoring and personalized healthcare, and revolutionize today’s healthcare system, especially in remote areas. The need for POC devices strongly drives the development of novel biosensor technology. Building a small, fast, simple, and sensitive platform for biomolecule detection is a challenge that relies on the integration of multiple fields of expertise and engineering. Optical nanoresonators have shown great promise as label-free biosensors because of direct light coupling and sub-wavelength sensing modes. Metallic nanoresonators with localized surface plasmon resonances (LSPR) are already well studied and were proven a solid alternative to the commercialized surface plasmon resonance (SPR) sensors. More recently, dielectric nanoresonators have also gained traction due to the reduced losses and the ability to manipulate both the electric and magnetic components of the incident light. In this thesis, we advance the field of biosensing and use optical nanoresonators as operative platforms relevant for disease diagnosis and treatment monitoring. By combining different optimized optical nanoresonators, both metallic and dielectric, with state-of-the-art microfluidics and surface chemistry, we have developed and tested several detection platforms. We first focused on developing a microfluidic lab-on-chip device for multiplexed biosensing utilizing the LSPR of gold nanoresonator arrays. By simultaneously tracking the extinction of 32 sensor arrays, we demonstrated multiplexed quantitative detection of four breast cancer markers in human serum. We showed that with well-optimized immunoassays, a low limit of detection (LOD) can be reached, paving the way towards clinically-relevant POC devices. Additionally, we implemented silicon nanoresonators supporting Mie resonances into functional and clinically-relevant applications. By integrating several arrays of Si nanoresonators with state-of-the-art microfluidics, we demonstrated their ability to detect cancer markers in human serum with high sensitivity and high specificity. Furthermore, we showed that the fabrication of Si nanoresonator array using low cost and scalable projection lithography leads to sufficiently low limits of detection, while enabling cheaper and faster sensor production for future POC applications. We also investigated the respective role of electric and magnetic dipole resonances and showed that they are associated with two different transduction mechanisms: resonance redshift and extinction decrease. Our work advances the development of future point-of-care sensing platforms for fast and low cost health monitoring at the molecular scale.La instrumentación Point-of-care (POC) es compacta, móvil y permite una detección rápida, razón por la que se prevé que sean de gran ayuda en áreas como el diagnostico precoz, la monitorización de tratamientos y la medicina personalizada, revolucionando los modelos sanitarios, especialmente en las zonas de difícil acceso y con menos recursos. La necesidad de este tipo de dispositivos impulsa el desarrollo de novedosas tecnologías en el campo de los bio-sensores. Diseñar equipos para la detección de bio-moléculas que sean rápidos, pequeños y sencillos es un reto que requiere la integración de múltiples campos de la ciencia y la ingeniería. Los nano-resonadores ópticos muestran un gran potencial como bio-sensores sin necesidad de marcaje, gracias a su capacidad de acoplase directamente con la luz en modos menores que la longitud de onda. Los nano-resonadores metálicos basados en resonancias plasmónicas superficiales localizadas (LSPR) han sido estudiados y han demostrado ser una firme alternativa a los ya comerciales basados en resonancias plasmónicas superficiales (SPR). Los nano-resonadores dieléctricos han sido recientemente objeto de atención debido a sus bajas perdidas y la capacidad de manipular los componentes eléctricos y magnéticos de la luz. En esta tesis presentamos avances en el campo de la bio-detección y en el uso de los nano-resonadores ópticos como potenciales herramientas para la detección de enfermedades y monitorización de los tratamientos. Hemos desarrollado y evaluado distintas plataformas de detección combinando los nano-resonadores ópticos, tanto metálicos como dieléctricos, con las más avanzadas técnicas de microfluídica y química de superficies. En primer lugar, nos centramos en el desarrollo de un dispositivo microfluídico basado en sensores LSPR de oro que permite multiplexar 32 canales. Los 32 sensores se monitorizan en tiempo real para demostrar la cuantificación de 4 marcadores de cáncer de mama en suero sanguíneo humano. Demostramos que mediante la optimización de los ensayos se pueden alcanzar bajos límites de detección (LOD), lo que allana el camino hacia dispositivos POC de uso clínico. Por otro lado, hemos utilizado los nano-resonadores de silicio integrados con la microfluídica para también detectar marcadores de cáncer en suero. Estos sensores, cuyo principio de funcionamiento se basa en resonancias de MIE, han demostrado ser una alternativa razonable a los sensores de oro. Además, demostramos que un proceso de fabricación de nano-resonadores de silicio rápido, escalable y de bajo coste da lugar a límites de detección suficientes para la producción de futuras POC. También realizamos un minucioso estudio del rol de las resonancias eléctricas y magnéticas en dichos sensores y su relación con el desplazamiento y el cambio magnitud de la resonancia del sensor global. Nuestro trabajo es un avance en el desarrollo de futuros instrumentos POC rápidos y baratos en el ámbito de la salud a escala molecular

Unravelling the Role of Electric and Magnetic Dipoles in Biosensing with Si Nanoresonators

High refractive index dielectric nanoresonators are attracting much attention due to their ability to control both electric and magnetic components of light. Combining confined modes with reduced absorption losses, they have recently been proposed as an alternative to nanoplasmonic biosensors. In this context, we study the use of semi-random silicon nanocylinder arrays, fabricated with simple and scalable colloidal lithography for the efficient and reliable detection of biomolecules in biological samples. Interestingly, electric and magnetic dipole resonances are associated to two different transduction mechanisms: extinction decrease and resonance redshift, respectively. By contrasting both observables, we identify clear advantages in tracking changes in the extinction magnitude. Our data demonstrate that, despite its simplicity, the proposed platform is able to detect prostate specific antigen (PSA) in human serum with limits of detection meeting clinical needs.Peer Reviewe

Incidentally Detected Kaposi Sarcoma of Adrenal Gland with Anaplastic Features in an HIV Negative Patient

Kaposi sarcoma (KS), a vascular tumor caused by infection with human herpesvirus 8 (HHV8), is a systemic disease that can present with cutaneous lesions with or without visceral involvement. Very few cases of KS, most of which were associated with AIDS, have been reported in the adrenal gland. Anaplastic transformation of KS is a rare clinical presentation known as an aggressive disease with local recurrence and metastatic potential. We report here a 47-year-old HIV negative male presented with extra-adrenal symptoms and had an incidentally detected anaplastic adrenal KS exhibited aggressive clinical course. To the best of our knowledge, this is the first case of anaplastic primary adrenal KS without mucocutaneous involvement but subsequently developed other side adrenal metastases in an HIV negative patient