CytoSensor system integration and design is driven by requirements generated by the need to complete biological experiment operations. The system is used for toxin-based detection which will identify and quantify unknown input toxins by using a biosensor based on a living fish chromatophore. The system consists of 3 main parts: biosensor, data acquisition and data interpretation. This thesis is focused on data acquisition. Acquisition, in this case, is via a color camera since the cells have an easily measurable visual output. The major initial task is to select the hardware specifications that satisfy user requirements. Components are obtained from different vendors. The understanding of each component is, therefore, very important to maximize the system performance and compatibility. The second major task is to design the software interface and components to manage the data acquisition. This can be separated into 2 parts. The first part is acquisition management and control. The second part is the human interface. This thesis focuses on the human interface. The human interface is the part that communicates between the user and the system. The system will send the system status to the user. The user will then direct the system through the operation. Operators may not be familiar with complicated computerized systems. A user-friendly interface is important to reduce mistakes and to facilitate the operation. The goal of this design is to direct the user from a single look at the interface. The interface should therefore contain all the useful and necessary information. The design of the user interface begins with gathering the necessary information and making a decision about which information is important to deliver to the user. A clean, tidy and informative user interface will lead to efficient operation. The design methodology is to group the same information within the same area and be consistent. Machine operation is very important, as well. In order to reduce the confusion in system operation, the machine operating protocol is designed to be very similar to the traditional protocol. Design of the machine operation is through interactions with the user. Sending user information to the machine will be handled by the system management program. By simulating the user scenario, each state change will lead to changing of the state of the machine, as well. The scenario is implemented in a state-like diagram. This state diagram must be implemented carefully in order to be able to handle all the cases and exceptions. The last and most important part is putting all the components together and testing the system. All possible scenarios and features listed before designing will be tested at this point. The last test is to run actual experiments with the system. After all the tests are satisfied, the system is delivered to the user. At this time, the user might give more feedback on the system. In conclusion, the overall goal of designing this system is not only to make the system for this specific application. However, the goal is to design a general application that will be able to apply to different sensor application. By changing the core management and hardware, the software can easily fit another sensor application. / Graduation date: 2003

The purpose of the thesis is to design and develop a network of automated, distributed, living cell-based sensors, called CytoSensors. Their main role is to detect a variety of biological and chemical toxins. The system is designed to help researchers to carry out multitude of experiments, in order to build a practical knowledge base in toxin detection. The network is developed in accordance with industry standards, to be used and deployed for prevention in inhospitable environments such as battlefields, toxic urban locations or polluted agricultural regions. The sensor is composed of a processing unit (processor and memory), an archiving unit (permanent data storage), a communication unit, input devices attached to a data acquisition unit, and control devices. The CytoSensor is specifically designed to acquire and analyze visual information about the living cells: hence cameras are used as input devices and frame grabbers are used as the digitizers. The control devices are additional external devices developed to help control and automate the process of data acquisition: they comprise light intensity control USB boards to provide the correct amount of light to view the cells, touch panels for user-instrument interaction, and bar code readers to identify vials and experiments. The software, on the other hand, is a complex mosaic of different elements, each of which has a specific task to accomplish. These building blocks include the real-time acquisition, archiving, networking, processing, modelling, sensor output presentation and user interfaces. Our goal is to develop, integrate and optimize all these components to produce a viable and working device. The prototypes evolved from an offline, portable sensor equipped with a single high-resolution CCD camera and high-quality optics, to distributed online sensors with multiplexed CCD cameras and affordable optics. The acquisition board digitizes in real time the images from one to twelve multiplexed high resolution cameras. Several operational requirements must be met. First, a fault-tolerant and stable control over the input devices and control devices must be provided. Secondly, acquisition timing errors should be minimized as a trade-off between performance and the use of a low-cost, general-purpose, industry-standard operating system such as Microsoft Windows NT. Finally, in order to reduce development time and increase code reusability, a common abstraction layer is designed to provide for flexible use with various types of digitizers and cameras. As part of a distributed detection network, each sensor is able to exchange data with other "trusted" sensors and users, and to allow remote control of certain tasks. The sensor may be seen as a node capable of transmitting and receiving acquired or processed data to a distant device (another sensor, a workstation or a PDA) for visualization, inspection and decision-making by a front-end user. Each node on the network provides a set of complementary services including data acquisition, data processing, communication and system. The mandatory system service monitors the local system performance and manages data archiving. The communication service connects the various services on the network by enabling message-passing, file transfer and caching. The sensor network integrates a lightweight, interoperable and flexible RPC (Remote Procedure Call) protocol to achieve real-time control and monitoring of these distributed resources. A reliable embedded database system is used to store metadata bound to acquired and processed images. This database is also used to maintain information on neighbor nodes, and to check access credentials of available local services. Finally, by adding store-and-forward messaging capabilities, the application can be extended to work in wireless and mobile networks. / Graduation date: 2003

To protect the environment and save human lives, the detection of various hazardous toxins of biological or chemical origin has been a major challenge to the researchers at Oregon State University. Living fish cells can indicate the presence of a wide range of toxins by reactions such as changing color and shape changes. A research team in Electrical and Computer Engineering Department is developing a hybrid detection device (Cytosensor) that combines biological reaction and digital technology. The functions of Cytosensor can be divided into three parts, which are real-time image acquisition, data processing and statistical data analysis. User-friendly Web-Based Distributed Applications (WBDA) for Cytosensor offer various utilities. WBDA allow the users to control and observe the local Cytosensor, search and retrieve data acquired by the sensor network, and process the acquired images remotely using only a web browser. Additionally, these applications minimize the user's exposure to dangerous chemicals or biological products. This thesis describes the design of a remote controller, system observer, remote processor, and search engine using JAVA applets, XML, Perl, MATLAB, and Peer-to-Peer models. Furthermore, the implementations of image segmentation technique in MATLAB and the Machine Vision Algorithm in JAVA for independent web-based processing are investigated. / Graduation date: 2003

<p>This thesis describes the use of a new technology, the Cytosensor^® microphysiometer, in the in vitro evaluation of cytotoxic drugs, using the lymphoma cell line U-937 GTB and primary cultures of tumour cells from patients as model systems. The method was specifically applied to study the metabolic effects of the novel cyanoguanidine N-(6-(4-chlorophenoxy)hexyl)-N’-cyano-N’’-4-pyridylguanidine, CHS 828, currently in phase I/II clinical trials. </p><p>The Cytosensor^® measures metabolic effects as changes in the rate of extracellular acidification of cells exposed to a drug by perfusion. A number of standard cytotoxic drugs were found to produce typical and reproducible acidification response patterns during observation times up to 20 h. There seemed to be a relationship between a decrease in acidification and cytotoxicity, measured in the fluorometric microculture cytotoxicity assay (FMCA), after 20-24 h of continuous drug exposure.</p><p>In U-937 cells, CHS 828 induced a cytotoxic effect characterised by a steep concentration-response relationship followed by a plateau. After 24 h of incubation the DNA and protein synthesis were turned off. CHS 828 was found to produce a rapid and prolonged increase in extracellular acidification and lactate production similar to that of the structurally related mitochondrial inhibitor m-iodobenzylguanidine (MIBG). The CHS 828 induced acidification was observed in cell lines as well as in cells from various tumour types from patients and probably originates from increased glycolytic flux. The effects may be secondary to block of oxidative phosphorylation in the mitochondria, but the relevance of the early acidification is not clear. CHS 828 seemed to induce a late, at approximately 15 h, inhibition of the glycolysis followed by loss of ATP and subsequent cell death. After exposure to MIBG the loss of ATP and cell death occurred earlier and in parallel. The effects of CHS 828 were not found to resemble those of the structurally related polyamine biosynthesis inhibitor methylglyoxal-bis(guanyl-hydrazone) (MGBG). Thus, CHS 828 may represent a new and, thus, interesting mode of cytotoxic action worthwhile for further development.</p><p>In combinatory studies, a synergistic interaction was demonstrated between CHS 828 and the non-toxic drug amiloride. Additive-to-synergistic effects were also seen between CHS 828 and the bioreductive cytotoxic drug mitomycin C. In U-937 cells as well as in tumour cells from patients, CHS 828 demonstrated synergistic interactions in combination with melphalan and etoposide. </p><p>It is concluded that measurement in the Cytosensor^® microphysiometer of early cellular metabolic changes is a feasible and potentially valuable complement to more conventional methods used in the evaluation of anticancer agents. </p>

This thesis describes the use of a new technology, the Cytosensor® microphysiometer, in the in vitro evaluation of cytotoxic drugs, using the lymphoma cell line U-937 GTB and primary cultures of tumour cells from patients as model systems. The method was specifically applied to study the metabolic effects of the novel cyanoguanidine N-(6-(4-chlorophenoxy)hexyl)-N’-cyano-N’’-4-pyridylguanidine, CHS 828, currently in phase I/II clinical trials. The Cytosensor® measures metabolic effects as changes in the rate of extracellular acidification of cells exposed to a drug by perfusion. A number of standard cytotoxic drugs were found to produce typical and reproducible acidification response patterns during observation times up to 20 h. There seemed to be a relationship between a decrease in acidification and cytotoxicity, measured in the fluorometric microculture cytotoxicity assay (FMCA), after 20-24 h of continuous drug exposure. In U-937 cells, CHS 828 induced a cytotoxic effect characterised by a steep concentration-response relationship followed by a plateau. After 24 h of incubation the DNA and protein synthesis were turned off. CHS 828 was found to produce a rapid and prolonged increase in extracellular acidification and lactate production similar to that of the structurally related mitochondrial inhibitor m-iodobenzylguanidine (MIBG). The CHS 828 induced acidification was observed in cell lines as well as in cells from various tumour types from patients and probably originates from increased glycolytic flux. The effects may be secondary to block of oxidative phosphorylation in the mitochondria, but the relevance of the early acidification is not clear. CHS 828 seemed to induce a late, at approximately 15 h, inhibition of the glycolysis followed by loss of ATP and subsequent cell death. After exposure to MIBG the loss of ATP and cell death occurred earlier and in parallel. The effects of CHS 828 were not found to resemble those of the structurally related polyamine biosynthesis inhibitor methylglyoxal-bis(guanyl-hydrazone) (MGBG). Thus, CHS 828 may represent a new and, thus, interesting mode of cytotoxic action worthwhile for further development. In combinatory studies, a synergistic interaction was demonstrated between CHS 828 and the non-toxic drug amiloride. Additive-to-synergistic effects were also seen between CHS 828 and the bioreductive cytotoxic drug mitomycin C. In U-937 cells as well as in tumour cells from patients, CHS 828 demonstrated synergistic interactions in combination with melphalan and etoposide. It is concluded that measurement in the Cytosensor® microphysiometer of early cellular metabolic changes is a feasible and potentially valuable complement to more conventional methods used in the evaluation of anticancer agents.