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A robotic manipulation system for automated selection and transfer of individual living cells of interest to analysis locations is presented. The system is the first in a series of operational components comprising an integrated system for the analysis of cell expression and metabolism at the single cell level. In our approach, we begin with a commonly used cell transfer technique, which uses glass...
In general, a formal specification for hardware-software integration is imperative for systematic design of automated systems. This specification requires a critical understanding of hardware and software systems at various levels of abstraction. The objective of the research we present is to develop a framework that serves as a specification for the implementation of an integrated system for the...
Major advances in the understanding in the biological sciences have been achieved by the precision automation of repetitive analytical procedures. The requirements for precision are pushed to the extreme when considering the analysis of cell function at the single cell level. Such analyses may include examination of gene expression, protein synthesis, and metabolic activity. The repetitive nature...
A novel system that has enabled the measurement of single-cell oxygen consumption rates is presented. The experimental apparatus includes a temperature controlled environmental chamber, an array of microwells etched in glass, and a lid actuator used to seal cells in the microwells. Each microwell contains an oxygen sensitive platinum phosphor sensor used to monitor the cellular metabolic rates. Custom...
Advancements in methods and algorithms for the measurement of oxygen consumption rates of single cells is presented. In this system a low density of randomly seeded eukaryotic cells are sealed in an array of microwells etched in glass (zero to three cells per microwell). The decrease in oxygen concentration inside each microwell in the array is measured yielding the oxygen consumption rates of the...
The analysis of cell function comprises an examination of gene expression, protein synthesis, and metabolic activity. In order to measure these parameters in single cells a means for signal transduction and amplification is required. Fluorescent molecules have been demonstrated to provide a powerful tool for this detection need when performing living cell analysis. The development of an image analysis...
PDMS (polydimethylsiloxane) is an important material for lab-on-a-chip, an emerging technology to automate biological and chemical operations in microfluidic devices. Laser manufacturing of materials suitable for use in laminate-based construction of lab-on-a-chip devices is an important step in the development of automated assembly line procedures. This study investigates the effect of laser cutting...
A proof-of-platform-concept architecture for the analysis of living cell arrays is presented. Initial experiments conducted show a conceptual design for single-cell multiparameter measurements in real time. As a first parameter, extensive development and experimentation have been conducted on a sensor that will measure single-cell oxygen consumption. Wafer-level processing for chip fabrication and...
The Microscale Life Sciences Center at the University of Washington is developing a suite of technologies that combine high-density cell assays and automatic platforms to achieve high-content living cell screening. The use of microfluidic cell arrays allows for the manipulation of cells and control of biochemical stimuli in parallel, making automated operation possible. However, screening the microscale...
Initial results of a comprehensive design software that optimizes parameters for fluorescence analysis of a user-defined fluorophore are presented. SLAP (spectral LED aggregation program) automatically selects configurations of LEDs that, in a fluorescence analysis system, maximize the emission signal (useful output) as a function of the excitation signal (interference), optics, photodetection modality,...
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