Electrical Characterization and Detection of Blood Cells and Stones in Urine

Urine contains an immense amount of information related to its physical, chemical, and biological components; hence, it is a promising tool in detecting various diseases. Available methods for detecting hematuria (blood in the urine) are not accurate. Results are influenced by many factors, such as, health and vitals of the patients, settings of the equipment and laboratories, which leads to false positive or false negative outputs. This necessitates the development of new, accurate, and easy-access methods that save time and effort. This study demonstrates a label-free and accurate method for detecting the presence of red and white blood cells (RBCs and WBCs) in urine by measuring the changes in the dielectric properties of urine upon increasing concentrations of both cell types. The current method could detect changes in the electrical properties of fresh urine over a short time interval, making this method suitable for detecting changes that cannot be recognized by conventional methods. Correcting these changes enabled the detection of a minimum cell concentration of 10² RBCs per ml which is not possible by conventional methods used in the labs except for the semi-quantitative method that can detect 50 RBCs per ml, but it is a lengthy and involved procedure, not suitable for high volume labs. This ability to detect a very small amount of both types of cells makes the proposed technique an attractive tool for detecting hematuria, the presence of which is indicative of problems in the excretory system. Furthermore, urolithiasis is also a very common problem worldwide, affecting adults, kids, and even animals. Calcium oxalate is the major constituent of urinary tract stones in individuals, primarily due to the consumption of high oxalate foods. The occurrence of urinary oxalate occurs by endogenous synthesis, especially in the upper urinary tract. In a normal, healthy individual, the excretion of oxalate ranges from 10 to 45 mg/day, depending on the age and gender, but the risk of stone formation starts at 25 mg/day depending on the health history of the individual. This study also addresses the detection of the presence of calcium oxalate in urine following the same label-free approach. This can be done by measuring the changes in the dielectric properties of urine with increasing concentrations of calcium oxalate hydrate (CaC₂O₄.H₂O). The current method could detect dynamic changes in the electrical properties of urine over a time interval in samples containing calcium oxalate hydrate even at a concentration as low as 10 μg/mL of urine, making this method suitable for detecting changes that cannot be recognized by conventional methods. The ability to detect a very small amount of stones makes it an attractive tool for detecting and quantifying stones in kidneys. Using a non-invasive method which also works as a precautionary measure for early detection of some severe ailments, holds a good scope. It forms the basis of the cytological examinations and molecular assays for the diagnosis of several diseases. This method can be considered a point-of-care test because the results can be instantaneously shared with the members of the medical team. Based on these results, it is anticipated that the present approach to be a starting point towards establishing the foundation for label-free electrical-based identification and quantification of an unlimited number of nano-sized particles

Battery-less near field communications (nfc) sensors for internet of things (iot) applications

L’ implementació de la tecnologia de comunicació de camp proper (NFC) en els telèfons intel·ligents no para de créixer degut a l’ús d’aquesta per fer pagaments, això, junt amb el fet de poder aprofitar l’energia generada pel mòbil no només per la comunicació, sinó també per transmetre energia, el baix cost dels xips NFC, i el fet de que els telèfons tinguin connectivitat amb internet, possibilita i fa molt interesant el disseny d’etiquetes sense bateria incorporant-hi sensors i poder enviar la informació al núvol, dins del creixent escenari de l’internet de les coses (IoT). La present Tesi estudia la viabilitat d’aquests sensors, analitzant la màxima distància entre lector i sensor per proveir la potència necessària, presenta tècniques per augmentar el rang d’operació, i analitza els efectes de certs materials quan aquests estan propers a les antenes. Diversos sensors han estat dissenyats i analitzats i son presentats en aquest treball. Aquests son: Una etiqueta que mesura la humitat de la terra, la temperatura i la humitat relativa de l’aire per controlar les condicions de plantes. Un sensor per detectar la humitat en bolquers, imprès en material flexible que s’adapta a la forma del bolquer. Dues aplicacions, una per estimació de pH i una altre per avaluar el grau de maduració de fruites, basats en un sensor de color. I, per últim, s’estudia la viabilitat de sensors en implants per aplicacions mèdiques, analitzant l’efecte del cos i proposant un sistema per augmentar la profunditat a la que aquests es poden llegir utilitzant un telèfon mòbil. Tots aquests sensors poden ser alimentats i llegits per qualsevol dispositiu que disposin de connexió NFC.La implementación de la tecnología de comunicaciones de campo cercano (NFC) en los teléfonos inteligentes no para de crecer debido al uso de esta para llevar a cabo pagos, esto, junto con el hecho de poder aprovechar la energía generada por el móvil no sólo para la comunicación, sino también para transmitir energía, el bajo coste de los chips NFC, i el hecho que los teléfonos tengan conectividad a internet, posibilita y hace muy interesante el diseño de etiquetas sin batería que incorporen sensores i poder enviar la información a la nube, enmarcado en el creciente escenario del internet de las cosas (IoT). La presente Tesis estudia la viabilidad de estos sensores, analizando la máxima distancia entre lector i sensor para proveer la potencia necesaria, presenta técnicas para aumentar el rango de operación, y analiza los efectos de ciertos materiales cuando estos están cerca de las antenas. Varios sensores han sido diseñados y analizados y son presentados en este trabajo. Estos son: Una etiqueta que mide la humedad de la tierra, la temperatura y la humedad relativa del aire para controlar las condiciones de plantas. Un sensor para detectar la humedad en pañales, impreso en material flexible que se adapta a la forma del pañal. Dos aplicaciones, una para estimación de pH y otra para evaluar el grado de maduración de frutas, basados en un sensor de color. Y, por último, se estudia la viabilidad de sensores en implantes para aplicaciones médicas, analizando el efecto del cuerpo y proponiendo un sistema para aumentar la profundidad a la que estos se pueden leer usando un teléfono móvil. Todos estos sensores pueden ser alimentados y leídos por cualquier dispositivo que disponga de conexión NFC.The implementation of near field communication (NFC) technology into smartphones grows rapidly due the use of this technology as a payment system. This, altogether with the fact that the energy generated by the phone can be used not only to communicate but for power transfer as well, the low-cost of the NFC chips, and the fact that the smartphones have connectivity to internet, makes possible and very interesting the design of battery-less sensing tags which information can be sent to the cloud, within the growing internet of things (IoT) scenario. This Thesis studies the feasibility of these sensors, analysing the maximum distance between reader and sensor to provide the necessary power, presents techniques to increase the range of operation, and analyses the effects of certain materials when they are near to the antennas. Several sensors have been designed and analysed and are presented in this work. These are: a tag that measures the soil moisture, the temperature and the relative humidity of the air to control the conditions of plants. A moisture sensor for diapers, printed on flexible material that adapts to the diaper shape. Two applications, one for pH estimation and another for assessing the degree of fruit ripening, based on a colour sensor. And finally, the feasibility of sensors in implants for medical applications is studied, analysing the effect of the body and proposing a system to increase the depth at which they can be read using a mobile phone. All of these sensors can be powered and read by any NFC enabled device

Bacterial sensors and controllers based on organic bioelectronics

Bacterial infections and contaminations are worldwide problems, leading to morbidity and mortality, food waste and economic losses in a variety of industries. The situation is aggravated by the increased occurrence of antibiotic-resistant strains, identified by the WHO as one of the biggest threats to development, food security and public health today. The solution to this problem is complex and requires efforts from several different layers of the society, and different disciplines. The knowledge about microbiology has greatly advanced in the last decades and several powerful methods were introduced. However, in most clinical microbiology laboratories, culture-based techniques are still standard practice, representing a bottleneck in the diagnostic workflow. In this thesis, we prototype novel methods to detect and identify bacteria, aiming to reduce the time and workload for future microbiology research and diagnostics. Furthermore, a new methodology is devised to evaluate antimicrobial surface properties for relevant high-touch surfaces. In Paper I, we investigated whether conducting polymers can be applied for label-free electrochemical detection of bacteria. Employing a poly(3,4-ethylenedioxythiophene): polystyrenesulfonate (PEDOT:PSS)-based two-electrode sensor we demonstrated that potentiometric detection and quantification of Salmonella Enteritidis is possible within 15 min, without any sample pre-treatment. We show that the reduction of PEDOT:PSS electrode occurs by low molecular weight species secreted by Salmonella Enteritidis. To evaluate the genericity of the sensor, several uropathogenic strains were tested and we found that they could all be detected using the sensor. In its current form, the sensor is a prototype, and we aim to improve its sensitivity and introduce specificity. Electroactivity was shown to be a rather common characteristic of bacteria and consequentially, electrochemical methods for detection and characterization of microbes are gaining momentum. We envision that this field will provide novel diagnostic devices but also contribute to discoveries in basic science. Luminescent conjugated oligothiophenes, called optotracers, have previously been applied in microbiology to visualize extracellular matrix components in biofilms of Salmonella and Escherichia coli. In Paper II, we investigated the use of optotracers for detection and visualization of Staphylococcus aureus (S. aureus). We show that the optotracer HS-167 selectively binds to Staphylococci and can be used for fluorometric detection and quantification of S. aureus, as well as for staining and visualization using confocal microscopy. HS-167 displays an on-switch of fluorescence upon binding and it does not affect bacterial growth, which enabled us to develop a high-throughput assay where the fluorescence was plotted against bacterial density, measured as an increase in turbidity. The resulting slope was a quantifiable variable that we employed to compare binding of HS-167 to different species and strains. Diverse approaches collectively pointed to the cell envelope as the target for HS-167 binding. Finally, we showed that binding is highly dependent on the environmental conditions and those can be adjusted to tune the selectivity of HS-167. To improve optotracer design for detection of S. aureus, a better insight into the structure- function relationship is needed. In Paper III, we set out to establish a structured approach to optotracer screening that would enable us to compare optotracer performance. As we compared a library of ten different optotracers, we identified the length to be positively correlated and the total negative charge to be negatively correlated with the ability to detect S. aureus. A balance between the two was necessary to achieve the highest signal while maintaining selectivity. Selected optotracers were added to S. aureus and visualized under the confocal microscope. All localized in the cell envelope of the bacterium, as was previously observed for HS-167 (Paper II). We foresee that further insight into the binding mechanism will enable targeted optotracer design, and together with optimized assay conditions, specific detection of different bacterial species. Copper is known to possess antimicrobial properties, yet studies have reported discrepant results on its efficacy, especially in the clinical settings. Disagreeing results were ascribed to the lack of a standardized approach to evaluate the antimicrobial properties of copper surfaces. In Paper IV, we establish a multifaceted approach to address the effect of human touch, which we simulate by applying artificial sweat, on surface corrosion and antimicrobial properties of copper. We found that artificial sweat accelerates corrosion, leading to changes in surface appearance and wettability. Corrosion did not negatively affect the antimicrobial properties of copper as these surfaces killed bacteria within minutes, regardless of ageing or corrosion product formation. The antimicrobial effect is ascribed in part to copper ions released from the surface and in part to direct surface contact. To further validate the results of this study, other bacterial species need to be tested. Since high touch surfaces are likely to collect a lot of microbes over time, it would be of interest to determine how the bacterial load affects the antimicrobial properties

Determination of antibiotic susceptibility of the bacteria causing urinary tract infections using a novel lab-on-a-chip design

Urinary tract infections (UTIs) are one of the most common types of bacterial infection in the UK, and also are expensive to treat costing the National Health Service ~£54 million between 2016 and 2017. Culture-based antibiotic susceptibility testing (AST) is used to identify an antibiotic to treat drug-resistant urinary tract infections and takes 48 hours to complete. Faster prescription of effective antibiotics should reduce the risk of sepsis and poor clinical outcomes. To address this need, we developed a Lab-on-a-Chip (LOC) based method to conduct electrochemical AST using screen-printed macroelectrodes (SPEs) and antibiotic-loaded hydrogels. SPEs were fabricated using carbon-graphite based inks, with resazurin bulk modified SPEs (R-SPEs) being fabricated through modification of the SPEs WE. Polyvinyl alcohol (PVA) based hydrogels were loaded with the following antibiotics were used; cephalexin, ceftriaxone, colistin, gentamicin, piperacillin, trimethoprim and vancomycin as well as an antibiotic-free control. LOC devices were then designed to encapsulate both the R-SPEs and the antibiotic hydrogels to enable multiplexed electrochemical AST to occur on a single device. In the initial testing of the R-SPEs and the antibiotic hydrogels independently of a LOC device, antibiotic susceptibility could be determined in 90 minutes for E. coli. After the preliminary work, eight chambered LOC devices were spiked with simulated UTI samples. Each chamber contained an R-SPE and an antibiotic hydrogel. After an incubation step, susceptibility of Escherichia coli and Klebsiella pneumoniae could be established in 85 minutes of testing which is significantly faster than the 48 hours required for conventional culture-based AST. The sensitive detection of resazurin afforded by using the electrochemical detection methodology incorporated onto a LOC device described here offers an inexpensive and simple method for the determination of antibiotic susceptibility that is faster than using a culture-based approach

Using wireless sensors and networks program for chemical particle propagation mapping and chemical source localization

Chemical source localization is a challenge for most of researchers. It has extensive applications, such as anti-terrorist military, Gas and oil industry, and environment engineering. This dissertation used wireless sensor and sensor networks to get chemical particle propagation mapping and chemical source localization. First, the chemical particle propagation mapping is built using interpolation and extrapolation methods. The interpolation method get the chemical particle path through the sensors, and the extrapolation method get the chemical particle beyond the sensors. Both of them compose of the mapping in the whole considered area. Second, the algorithm of sensor fusion is proposed. It smooths the chemical particle paths through integration of more sensors\u27 value and updating the parameters. The updated parameters are associated with including sensor fusion among chemical sensors and wind sensors at same positions and sensor fusion among sensors at different positions. This algorithm improves the accuracy and efficiency of chemical particle mapping. Last, the reasoning system is implemented aiming to detect the chemical source in the considered region where the chemical particle propagation mapping has been finished. This control scheme dynamically analyzes the data from the sensors and guide us to find the goal. In this dissertation, the novel algorithm of modelling chemical propagation is programmed using Matlab. Comparing the results from computational fluid dynamics (CFD) software COMSOL, this algorithm have the same level of accuracy. However, it saves more computational times and memories. The simulation and experiment of detecting chemical source in an indoor environment and outdoor environment are finished in this dissertation --Abstract, page iii

Navigation system based in motion tracking sensor for percutaneous renal access

Tese de Doutoramento em Engenharia BiomédicaMinimally-invasive kidney interventions are daily performed to diagnose and treat several renal diseases. Percutaneous renal access (PRA) is an essential but challenging stage for most of these procedures, since its outcome is directly linked to the physician’s ability to precisely visualize and reach the anatomical target. Nowadays, PRA is always guided with medical imaging assistance, most frequently using X-ray based imaging (e.g. fluoroscopy). Thus, radiation on the surgical theater represents a major risk to the medical team, where its exclusion from PRA has a direct impact diminishing the dose exposure on both patients and physicians. To solve the referred problems this thesis aims to develop a new hardware/software framework to intuitively and safely guide the surgeon during PRA planning and puncturing. In terms of surgical planning, a set of methodologies were developed to increase the certainty of reaching a specific target inside the kidney. The most relevant abdominal structures for PRA were automatically clustered into different 3D volumes. For that, primitive volumes were merged as a local optimization problem using the minimum description length principle and image statistical properties. A multi-volume Ray Cast method was then used to highlight each segmented volume. Results show that it is possible to detect all abdominal structures surrounding the kidney, with the ability to correctly estimate a virtual trajectory. Concerning the percutaneous puncturing stage, either an electromagnetic or optical solution were developed and tested in multiple in vitro, in vivo and ex vivo trials. The optical tracking solution aids in establishing the desired puncture site and choosing the best virtual puncture trajectory. However, this system required a line of sight to different optical markers placed at the needle base, limiting the accuracy when tracking inside the human body. Results show that the needle tip can deflect from its initial straight line trajectory with an error higher than 3 mm. Moreover, a complex registration procedure and initial setup is needed. On the other hand, a real-time electromagnetic tracking was developed. Hereto, a catheter was inserted trans-urethrally towards the renal target. This catheter has a position and orientation electromagnetic sensor on its tip that function as a real-time target locator. Then, a needle integrating a similar sensor is used. From the data provided by both sensors, one computes a virtual puncture trajectory, which is displayed in a 3D visualization software. In vivo tests showed a median renal and ureteral puncture times of 19 and 51 seconds, respectively (range 14 to 45 and 45 to 67 seconds). Such results represent a puncture time improvement between 75% and 85% when comparing to state of the art methods. 3D sound and vibrotactile feedback were also developed to provide additional information about the needle orientation. By using these kind of feedback, it was verified that the surgeon tends to follow a virtual puncture trajectory with a reduced amount of deviations from the ideal trajectory, being able to anticipate any movement even without looking to a monitor. Best results show that 3D sound sources were correctly identified 79.2 ± 8.1% of times with an average angulation error of 10.4º degrees. Vibration sources were accurately identified 91.1 ± 3.6% of times with an average angulation error of 8.0º degrees. Additionally to the EMT framework, three circular ultrasound transducers were built with a needle working channel. One explored different manufacture fabrication setups in terms of the piezoelectric materials, transducer construction, single vs. multi array configurations, backing and matching material design. The A-scan signals retrieved from each transducer were filtered and processed to automatically detect reflected echoes and to alert the surgeon when undesirable anatomical structures are in between the puncture path. The transducers were mapped in a water tank and tested in a study involving 45 phantoms. Results showed that the beam cross-sectional area oscillates around the ceramics radius and it was possible to automatically detect echo signals in phantoms with length higher than 80 mm. Hereupon, it is expected that the introduction of the proposed system on the PRA procedure, will allow to guide the surgeon through the optimal path towards the precise kidney target, increasing surgeon’s confidence and reducing complications (e.g. organ perforation) during PRA. Moreover, the developed framework has the potential to make the PRA free of radiation for both patient and surgeon and to broad the use of PRA to less specialized surgeons.Intervenções renais minimamente invasivas são realizadas diariamente para o tratamento e diagnóstico de várias doenças renais. O acesso renal percutâneo (ARP) é uma etapa essencial e desafiante na maior parte destes procedimentos. O seu resultado encontra-se diretamente relacionado com a capacidade do cirurgião visualizar e atingir com precisão o alvo anatómico. Hoje em dia, o ARP é sempre guiado com recurso a sistemas imagiológicos, na maior parte das vezes baseados em raios-X (p.e. a fluoroscopia). A radiação destes sistemas nas salas cirúrgicas representa um grande risco para a equipa médica, aonde a sua remoção levará a um impacto direto na diminuição da dose exposta aos pacientes e cirurgiões. De modo a resolver os problemas existentes, esta tese tem como objetivo o desenvolvimento de uma framework de hardware/software que permita, de forma intuitiva e segura, guiar o cirurgião durante o planeamento e punção do ARP. Em termos de planeamento, foi desenvolvido um conjunto de metodologias de modo a aumentar a eficácia com que o alvo anatómico é alcançado. As estruturas abdominais mais relevantes para o procedimento de ARP, foram automaticamente agrupadas em volumes 3D, através de um problema de optimização global com base no princípio de “minimum description length” e propriedades estatísticas da imagem. Por fim, um procedimento de Ray Cast, com múltiplas funções de transferência, foi utilizado para enfatizar as estruturas segmentadas. Os resultados mostram que é possível detetar todas as estruturas abdominais envolventes ao rim, com a capacidade para estimar corretamente uma trajetória virtual. No que diz respeito à fase de punção percutânea, foram testadas duas soluções de deteção de movimento (ótica e eletromagnética) em múltiplos ensaios in vitro, in vivo e ex vivo. A solução baseada em sensores óticos ajudou no cálculo do melhor ponto de punção e na definição da melhor trajetória a seguir. Contudo, este sistema necessita de uma linha de visão com diferentes marcadores óticos acoplados à base da agulha, limitando a precisão com que a agulha é detetada no interior do corpo humano. Os resultados indicam que a agulha pode sofrer deflexões à medida que vai sendo inserida, com erros superiores a 3 mm. Por outro lado, foi desenvolvida e testada uma solução com base em sensores eletromagnéticos. Para tal, um cateter que integra um sensor de posição e orientação na sua ponta, foi colocado por via trans-uretral junto do alvo renal. De seguida, uma agulha, integrando um sensor semelhante, é utilizada para a punção percutânea. A partir da diferença espacial de ambos os sensores, é possível gerar uma trajetória de punção virtual. A mediana do tempo necessário para puncionar o rim e ureter, segundo esta trajetória, foi de 19 e 51 segundos, respetivamente (variações de 14 a 45 e 45 a 67 segundos). Estes resultados representam uma melhoria do tempo de punção entre 75% e 85%, quando comparados com o estado da arte dos métodos atuais. Além do feedback visual, som 3D e feedback vibratório foram explorados de modo a fornecer informações complementares da posição da agulha. Verificou-se que com este tipo de feedback, o cirurgião tende a seguir uma trajetória de punção com desvios mínimos, sendo igualmente capaz de antecipar qualquer movimento, mesmo sem olhar para o monitor. Fontes de som e vibração podem ser corretamente detetadas em 79,2 ± 8,1% e 91,1 ± 3,6%, com erros médios de angulação de 10.4º e 8.0 graus, respetivamente. Adicionalmente ao sistema de navegação, foram também produzidos três transdutores de ultrassom circulares com um canal de trabalho para a agulha. Para tal, foram exploradas diferentes configurações de fabricação em termos de materiais piezoelétricos, transdutores multi-array ou singulares e espessura/material de layers de suporte. Os sinais originados em cada transdutor foram filtrados e processados de modo a detetar de forma automática os ecos refletidos, e assim, alertar o cirurgião quando existem variações anatómicas ao longo do caminho de punção. Os transdutores foram mapeados num tanque de água e testados em 45 phantoms. Os resultados mostraram que o feixe de área em corte transversal oscila em torno do raio de cerâmica, e que os ecos refletidos são detetados em phantoms com comprimentos superiores a 80 mm. Desta forma, é expectável que a introdução deste novo sistema a nível do ARP permitirá conduzir o cirurgião ao longo do caminho de punção ideal, aumentado a confiança do cirurgião e reduzindo possíveis complicações (p.e. a perfuração dos órgãos). Além disso, de realçar que este sistema apresenta o potencial de tornar o ARP livre de radiação e alarga-lo a cirurgiões menos especializados.The present work was only possible thanks to the support by the Portuguese Science and Technology Foundation through the PhD grant with reference SFRH/BD/74276/2010 funded by FCT/MEC (PIDDAC) and by Fundo Europeu de Desenvolvimento Regional (FEDER), Programa COMPETE - Programa Operacional Factores de Competitividade (POFC) do QREN

MEMS Technology for Biomedical Imaging Applications

Biomedical imaging is the key technique and process to create informative images of the human body or other organic structures for clinical purposes or medical science. Micro-electro-mechanical systems (MEMS) technology has demonstrated enormous potential in biomedical imaging applications due to its outstanding advantages of, for instance, miniaturization, high speed, higher resolution, and convenience of batch fabrication. There are many advancements and breakthroughs developing in the academic community, and there are a few challenges raised accordingly upon the designs, structures, fabrication, integration, and applications of MEMS for all kinds of biomedical imaging. This Special Issue aims to collate and showcase research papers, short commutations, perspectives, and insightful review articles from esteemed colleagues that demonstrate: (1) original works on the topic of MEMS components or devices based on various kinds of mechanisms for biomedical imaging; and (2) new developments and potentials of applying MEMS technology of any kind in biomedical imaging. The objective of this special session is to provide insightful information regarding the technological advancements for the researchers in the community