Polymerase Chain Reaction (PCR) detection paves the way to reliable and rapid diagnosis of diseases and has been used extensively since its introduction. Many miniaturized PCR systems were presented by microfluidics and lab-on-a-chip community. However, most of the developed systems did not employ real-time detection and thus required post-PCR processes to obtain results. Among the few real-time PCR systems, almost all of them aimed for medical applications and those for plant pathogen diagnosis systems are almost non-existent in the literature. In this work, we are presenting a portable system that employs microfluidics PCR system with integrated optical systems to accomplish real-time quantitative PCR for plant pathogen diagnosis. The system is comprised of a PCR chip that has a chamber for PCR sample with integrated metal heaters fabricated by standard microfabrication procedures, an optical system that includes lenses, filters, a dichroic mirror and a photomultiplier tube (PMT) to achieve sensitive fluorescence measurement capability and a computer control system for Proportional Integral Derivative (PID) control and data acquisition. The optical detection system employs portable components and has a size of 3.9 x 5.9 x 11.9 cm which makes it possible to be used in field settings. On the device side, two different designs are used. The first design includes a single chamber in a 25.4 x 25.4 mm device and the capacity of the chamber is 9 micro-liters which is sufficient to do gel electrophoresis verification. The second design has three 2.2 micro-liter chambers squeezed in the same size device while having smaller volume to increase high throughput of the system. The operation of the system was demonstrated using Fusarium oxysporum spf. lycopersici which is a fungal plant pathogen that affects crops in the USA. In the presence of the plant pathogen, noticeable increases in the photomultiplier tube output were observed which means successful amplifications and detections occurred. The results were confirmed using gel electrophoresis which is a conventional post-PCR process to determine the existence and length of the amplified DNA. Clear bands located in the expected position were observed following the gel electrophoresis. Overall, we have presented a portable PCR system that has the capability of detecting plant pathogens.

El objetivo del presente trabajo es demostrar la viabilidad de un negocio orientado al rastreo satelital de mascotas a través de un microchip insertado en los canes, el cual se encuentra ligado a un aplicativo móvil que ofrece servicios adicionales como la ubicación de veterinarias y pet shops aledaños, así como un historial de paseadores de perros calificados. Para ello, se detalla el planeamiento estratégico que incluye un análisis interno y externo de la organización. Asimismo, se explican los resultados de la investigación de mercado. También, se precisan los planes de marketing, operaciones, económico-financiero y la estructura organizacional, junto con la gestión de recursos humanos. En la investigación, se utiliza tanto la metodología cualitativa como la cuantitativa. Los resultados indican la viabilidad del negocio, así como su aceptación por parte del público objetivo y su rentabilidad para los accionistas e inversionistas de la empresa. Se concluye, por lo tanto, que el modelo de negocio de GPS PETS es viable, alcanzable y expandible en el mercado peruano. / The aim of this project is to prove the viability of a business created to track pets through a microchip inserted in dogs, which is linked to a mobile application that also offers certain services such as the location of pet shops and vet centers nearby. It also contains a list of dog walkers who are highly qualified. Therefore, a strategic planning is listed, which includes an internal and external analysis of the organization. Plus, the results of the market research are explained in detail. Moreover, the marketing, economic, and operational plans are described along with the organizational structure and the human resources management. In this investigation, the quantitative and qualitative research methods are used. The results indicate the viability of this business, as well as its acceptance by the target market, and its profitability for the company’s shareholders and investors. To sum up, it is concluded that GPS PETS business model is achievable, sustainable and expandable in the Peruvian market. / Trabajo de investigación

My work describes design of the universal development board usefull with a various types of microcontrollers and the unified interfaces for the microcontroller's module board. It introduce the design and the interface and list of the Microchip PIC microcontrollers which can be used with development board. It also describes the features of the PIC18F4550 microcontroller and means about an different types of anothers manufactures. There is also describe of the design, creating the schemes of the boards, printed circuit boards and 3D visualization models in my work.

<p>Routing techniques for plain ’flat’ microchips have been developed extensively and will soon reach its limitations. One natural step would be to develop chips which are manufactured in a more cubic type of volume, as oppose to the classical flat design. </p><p>This thesis proposes a method for routing two-point nets across a three- dimensional channel. The height required by this algorithm is of the order <i>O(n</i> ^(3/2)), where n is the number of terminals on a square top-layer with the side <i>2 (n</i>^(1/2)). </p><p>The algorithm proposed here is based on"On Routing Two-Point Nets Across a Channel", by Ron Y. Pinter [9], and the concepts from this paper are explainedin this thesis to familiarise the reader these. </p><p>It is also shown that the proposed algorithm is more effective in its volume than the two-dimensional counterpart. The algorithm here is of the order <i>O(n</i>^(3/2)) with the two-dimensional algorithm of the order <i>O</i>(<i>n</i>²).</p>

Detection of cellular messenger RNA is a useful diagnostic strategy for the detection of patho-gens. A rapid and sensitive method for on-site detection of specific pathogens would be of great use in a number of fields. For example, a simple and inexpensive method for the detection of harmful biological agents in train stations and airports is useful for national security. Rapid detection of pathogenic E. coli strains in food production would also be of great benefit in ensuring the safety and quality of our food supply. Here we present a method for the rapid de-tection of cellular mRNA. This system is based on the 3’-labeling approach in which targeted RNA is simultaneously extended and labeled with the use of biotin labeled-dNTPs and DNA po-lymerase on an immobilized nucleic acid probe. The biotin is subsequently converted to an enzymatic label, which produces a detectable chemiluminescent reaction in the presence of substrate. Detection time of this system is short (approximately 20 minutes) because there is no need for amplification by PCR, transcription, or fluorophore labeling. This novel methodology has been successfully demonstrated by selective detection of lac Z mRNA in a total RNA sample from E. coli.

Routing techniques for plain ’flat’ microchips have been developed extensively and will soon reach its limitations. One natural step would be to develop chips which are manufactured in a more cubic type of volume, as oppose to the classical flat design. This thesis proposes a method for routing two-point nets across a three- dimensional channel. The height required by this algorithm is of the order O(n (3/2)), where n is the number of terminals on a square top-layer with the side 2 (n(1/2)). The algorithm proposed here is based on"On Routing Two-Point Nets Across a Channel", by Ron Y. Pinter [9], and the concepts from this paper are explainedin this thesis to familiarise the reader these. It is also shown that the proposed algorithm is more effective in its volume than the two-dimensional counterpart. The algorithm here is of the order O(n(3/2)) with the two-dimensional algorithm of the order O(n2).

DNA gel electrophoresis is a critical analytical step in a wide spectrum of genomic analysis assays. Great efforts have been directed to the development of miniaturized microfluidic systems (“lab-on-a-chip” systems) to perform low-cost, high-throughput DNA gel electrophoresis. However, further progress toward dramatic improvements of separation performance over ultra-short distances requires a much more detailed understanding of the physics of DNA migration in the sieving gel matrix than is currently available in literature. The ultimate goal would be the ability to quantitatively determine the achievable level of separation performance by direct measurements of fundamental parameters (mobility, diffusion, and dispersion coefficients) associated with the gel matrix instead of the traditional trial-and-error process. We successfully established this predicting capability by measuring these fundamental parameters on a conventional slab gel DNA sequencer. However, it is difficult to carry out fast and extensive measurements of these parameters on a conventional gel electrophoresis system using single-point detection (2,000 hours on the slab gel DNA sequencer we used). To address this issue, we designed and built a new automated whole-gel scanning detection system for a systematic investigation of these governing parameters on a microfluidic gel electrophoresis device with integrated on-chip electrodes, heaters, and temperature sensors. With this system, we can observe the progress of DNA separation along the whole microchannel with high temporal and spatial accuracy in nearly real time. This is in contrast to both conventional slab gel imaging where the entire gel can be monitored, but only at one time frame after completion of the separation, and capillary electrophoresis systems that allows detection as a function of time, but only at a single detection location. With this system, a complete set of mobility, diffusion, and dispersion data can be collected within one hour instead of days or even months of work on a conventional sequencer under the same experimental conditions. The ability to acquire both spatial and temporal data simultaneously provides a more detailed picture of the separation process that can potentially be used to refine theoretical models and improve separation performance over ultra-short distances for the nextgeneration of electrophoresis technology.

<p>Promising applications in the design of various biological systems hold critical implications as heralded in the rising field of synthetic biology. But, to achieve these goals, the ability to synthesize in situ DNA constructs of any size or sequence rapidly, accurately and economically is crucial. Today, the process of DNA oligonucleotide synthesis has been automated but the overall development of gene and genome synthesis technology has far lagged behind that of gene and genome sequencing. This has meant that numerous ideas go unfulfilled due to scale, cost and impediments in the quality of DNA due to synthesis errors. </p><p>This thesis presents the development of a multi-tool ensemble platform targeted at gene synthesis. An inkjet oligonucleotide synthesizer is constructed to synthesize DNA microarrays onto silica functionalized cylic olefin copolymer substrates. The arrays are married to microfluidic wells that provide a chamber to for enzymatic amplification and assembly of the DNA from the microarrays into a larger construct. Harvested product is then amplified off-chip and error corrected using a mismatch endonuclease-based reaction. This platform has the potential to be particularly low-cost since it employs standard phosphoramidite reagents and parts that are cheaper than optical and electrochemical systems. Genes sized 160 bp to 993 bp were successfully harvested and, after error correction, achieved up to 94% of intended functionality.</p> / Dissertation

This thesis focuses on implementation of the swing-up, switching and stabilizing controllers for the rotary inverted pendulum. An energy based method to swing-up the pendulum and a state feedback controller to keep the pendulum in the upright position are employed. The mixed H2/H∞; state feedback controller is used to stabilize the pendulum with reduced oscillations. The results have been compared with the standard full state feedback and LQR. The Quanser rotary inverted pendulum is used as the testbed. All controllers are implemented in real-time using dSPACE 1104 rapid prototyping system. Microstick II with dsPIC33FJ128MC802 and Simulink embedded target for Microchip® is used as a standalone way to implement the controllers.

Kyoto University (京都大学) / 0048 / 新制・課程博士 / 博士(工学) / 甲第16864号 / 工博第3585号 / 新制||工||1542(附属図書館) / 29539 / 京都大学大学院工学研究科材料化学専攻 / (主査)教授 大塚 浩二, 教授 松原 誠二郎, 教授 田中 勝久 / 学位規則第4条第1項該当