Flow Cytometry: Digital Processing of Cellular Information

该项目包括进一步设计、建造和评估与流式细胞仪接口的数据采集和分析硬件。该硬件采用数字脉冲波形处理，在前期资助期间，我们开发并评估了第一代设计。在这项提案中，我们的目标是开发第二代硬件，以提供更高的细胞分析速度。我们还建议对在流式细胞术中使用数字脉冲处理可以获得的生物信息的类型以及提取这些信息所需的算法进行彻底的实践和理论检查。最后，我们提出是否可以通过提高流式细胞仪光学系统的空间分辨率来提高硬件的性能。拟议研究的预期意义依赖于观察到的是，该仪器在能力上是新颖的，并且可以提供使用其他仪器无法获得的细胞信息。第二代仪器的开发将使数字脉冲处理在流式细胞仪和细胞分选中完全实现。根据新的光学参数快速分析和分类细胞的能力应该会极大地扩展我们探索控制活细胞运行的细胞和分子因素的能力。这应该有助于更深入地了解细胞的正常生长和发育，以及异常生长和疾病状态。这可能会导致具有更好的诊断和预后能力的器械。我们相信，这个项目所必需的跨学科方法结合了非常不同的科学领域(工程学、光学和生物学)的专业知识，具有相当大的潜力来生产仪器和方法，从而显著扩展我们对活细胞结构和功能的理解。

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.