High-throughput vibrational cytometry

描述（由申请人提供）：流式细胞术是一种允许测量单个细胞或颗粒的各种特征的技术，通过观察它们在液体流中流动时的特性来确定。高速度的流动和大量的对象被分析强加了一些严格的标准上的方法可以用于分析。大多数已知的商业仪器目前使用光散射进行颗粒尺寸测量和荧光检测进行化学识别。然而，振动光谱是唯一的非侵入式光谱工具，它已被证明可以提供有关被询问样品的化学特定信息。假设基于非线性拉曼散射的振动光谱可以通过提供流动材料的快速和准确的化学识别来用作细胞计数的分析工具。在拟议的探索性（R21）研究中，化学物质的超快速分析的想法将进行实验测试，着眼于开发的仪器到商业流式细胞仪的潜在适应性。将建造一种新的仪器（目标1），以获得快速（即每秒10，000次分析）检测和检查活细胞所需的理想参数。将对所开发的仪器进行细致的测试（目标2），以确定其（a）灵敏度、（B）再现性、（c）信噪比和（e）光谱采集和分析速度。最后（目标3），开发和优化的仪器将使用活细胞模型系统进行测试，以了解振动光谱对细胞系统流式细胞术的适用性和潜在的权衡。该提案的长期目标是开发一种快速、非侵入性的细胞和颗粒分析方法，可应用于细胞/颗粒分析和分选、细菌病原体检测以及细胞与药物的相互作用。该仪器有望成为基础和临床生物医学研究的重要工具，并将通过提供一种以前所未有的速度水平分析细胞和化学结构的新方法来影响生物医学科学的许多领域。 公共卫生相关性：流式细胞术是一种用于对流体流中的微观颗粒进行计数、检查和分选的技术。它允许同时对单个细胞和颗粒的物理和/或化学特性进行多参数分析。一个真正的非侵入性流式细胞仪系统，能够提供化学特定的信息，提出并将开发和验证。所提出的技术在许多领域中具有应用，例如病理学、免疫学、毒理学和药理学。

DESCRIPTION (provided by applicant): Flow cytometry is a technology that allows a single cell or particle to be measured for a variety of characteristics, determined by looking at their properties while they flow in a liquid stream. High speed of flow and huge number of objects to be analyzed imposes some strict criteria on which methods can be used for analysis. Most of the known commercial instruments are currently using light scattering for particle sizing and fluorescence detection for chemical recognition. However, vibrational spectroscopy is the only non-invasive optical spectroscopy tool, which has proven to provide chemically-specific information about the interrogated sample. It is hypothesized that vibrational spectroscopy, based on nonlinear Raman scattering can be used to serve as an analytical tool for cytometry by providing rapid and accurate chemical recognition of flowing materials. In the proposed exploratory (R21) research, the idea of ultra-rapid analysis of chemical species will be experimentally tested with an eye on potential accommodation of the developed instrumentation to a commercial flow cytometer. A new instrument will be constructed (Aim 1) to attain the desired parameters needed for rapid, i.e. of the order of 10,000 analyses per second, detection and examination of living cells. The developed instrumentation will be meticulously tested (Aim 2) for its (a) sensitivity, (b) reproducibility, (c) signal- noise-ratio, and (e) speed of spectra acquisition and analysis. Finally (Aim 3), the developed and optimized instrumentation will be tested using a living cell model system to understand the applicability and potential trade-offs of vibrational spectroscopy to flow cytometry of cellular systems. The long-term of this proposal is to develop a rapid, non-invasive methodology for cell and particle analysis, which can be applied for cell/particle analysis and sorting, detection of bacterial pathogens, and cell interactions with pharmaceuticals. The proposed instrument is envisioned to become an important tool in fundamental and clinical biomedical research and will impact many areas of biomedical sciences by providing a novel way to analyze cells and chemical structures at unprecedented speed levels. PUBLIC HEALTH RELEVANCE: Flow cytometry is a technique for counting, examining, and sorting microscopic particles in a stream of fluid. It allows simultaneous multiparametric analysis of the physical and/or chemical characteristics of single cells and particles. A truly non-invasive flow cytometer system, capable of providing chemically specific information, is proposed and will be developed and validated. The proposed technology has applications in a number of fields, such as pathology, immunology, toxicology, and pharmacology.