Flow Cytometry: Digital Processing of Cellular Information

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This project involves the further design, construction, and evaluation of data acquisition and analysis hardware to be interfaced to a flow cytometer. This hardware employs digital pulse waveform processing, and during the prior period of funding, we developed and evaluated a first-generation design. In this proposal, we aim to develop second-generation hardware, that will provide improved rates of cellular analysis. We also propose a thorough practical and theoretical examination of the types of biological information that can be obtained using digital pulse processing in flow cytometry, and the algorithms that are required to extract this information. We finally propose to see whether we can improve the performance of the hardware through increasing the spatial resolution of the optics of the flow cytometer. The expected significance of the proposed research relies on the observation that the instrumentation is novel in capability, and can provide information about cells that is inaccessible using other instruments. The development of a second-generation instrument will allow full implementation of digital pulse processing in flow cytometry and cell sorting. The ability to rapidly analyze and sort cells according to novel optical parameters should greatly extend our ability to probe the cellular and molecular factors that govern the operation of living cells. This should facilitate a deeper understanding of normal cellular growth and development, as well as of abnormal growth and disease states. This might lead to instrumentation with improved diagnostic and prognostic capabilities. We are convinced that the interdisciplinary approach integral to this project, which combines expertise in very different scientific fields (engineering, optics, and biology), has considerable potential to produce instrumentation and methods that will significantly extend our understanding of the structure and function of living cells.