High-throughput vibrational cytometry

描述(申请人提供)：流式细胞术是一种技术，它允许测量单个细胞或颗粒的各种特性，通过观察它们在液体中流动时的特性来确定。高速的流动速度和巨大的待分析对象数量对分析方法提出了一些严格的标准。目前，大多数已知的商用仪器都使用光散射来测量颗粒尺寸，使用荧光检测来进行化学识别。然而，振动光谱学是唯一一种非侵入性的光学光谱学工具，它已被证明可以提供关于被询问样品的特定化学信息。基于非线性拉曼散射的振动光谱通过提供流动物质的快速和准确的化学识别，可以被用作细胞学的分析工具。在拟议的探索性研究(R21)中，将对化学物种的超快速分析的想法进行实验测试，并着眼于所开发的仪器对商业流式细胞仪的潜在适应性。将建造一台新仪器(目标1)，以获得快速检测活细胞所需的所需参数，即每秒10,000次分析的数量级。将仔细测试(目标2)所开发仪器的(A)灵敏度、(B)重现性、(C)信噪比和(E)光谱采集和分析的速度。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，以了解振动光谱在细胞系统的流式细胞术中的适用性和潜在的权衡。这项提议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，可用于细胞/颗粒分析和分选、细菌病原体的检测以及细胞与药物的相互作用。拟议的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分析细胞和化学结构的新方法，影响生物医学的许多领域。 与公共卫生相关：流式细胞术是一种对液体中的微小颗粒进行计数、检查和分类的技术。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。一种真正的非侵入性流式细胞仪系统被提出，并将被开发和验证，该系统能够提供化学特异性信息。这项拟议的技术在许多领域都有应用，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.