Eliminating Interference from Autofluorescence in Flow Cytometry

消除流式细胞术中自发荧光的干扰

• 批准号: 10484468
• 负责人: Giacomo Vacca
• 金额: $ 144.18万
• 依托单位: KINETIC RIVER CORPORATION
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10484468
• 关键词: Acquired Immunodeficiency Syndrome; Acute Myelocytic Leukemia; Algorithmic Analysis; Algorithmic Software; Algorithms; Antigens; Benchmarking; Biological; Biological Assay; Biomedical Research; Cancer Biology; Cell Count; Cell Membrane Permeability; Cells; Clinical; Collection; Color; Colorado; Communicable Diseases; Data; Data Analyses; Detection; Diagnosis; Diagnostic; Discipline; Dyes; Event; Excision; Fiber; Flow Cytometry; Fluorescence; Freeze Drying; Funding; Grant; Human; Immunologist; Immunology; Immunophenotyping; Kinetics; Label; Laboratories; Lasers; Lead; Lymphocyte; Malignant Neoplasms; Measures; Methods; Microbiology; Microspheres; Molecular Biology; Monitor; Myeloid Cells; NADH; Oncology; Output; Oxidation-Reduction; Patients; Phase; Ploidies; Research; Research Personnel; Rivers; Sampling; Shapes; Signal Transduction; Small Business Innovation Research Grant; Source; Speed; Stains; Surface; System; Technology; Testing; Time; Tumor Biology; United States National Institutes of Health; Viola; analog; cancer immunotherapy; clinically relevant; commercialization; computerized data processing; data acquisition; data handling; data quality; design; drug development; experimental study; fluorophore; improved; instrument; novel; optical spectra; programs; prototype; receptor; response; signal processing; stem cell therapy; time resolved data; time use; tool

PROJECT SUMMARY/ABSTRACT Flow cytometry (FC) is a powerful cell analysis tool relying on fluorescently labeling of cells. It can provide information on cell count, shape, size, DNA content, redox state, membrane permeability, and surface receptors, among other features. FC is an essential tool in research and clinical fields for tumor biology research, cancer and AIDS diagnostics, stem cell therapy, immunophenotyping, and cancer immunotherapy. One of the leading limitations of FC is that exogenously added fluorophores are not the only source of fluorescence in a sample. Autofluorescence (AF) can be, in some circumstances, a significant contributor to the signal present in each of the detection channels, often overlapping with the emission spectra. AF limits assay sensitivity and causes problems in data interpretation, which can lead to erroneous scientific conclusions. While several approaches have been employed to eliminate AF, there are no satisfactory solutions available. Kinetic River has been developing novel methods utilizing fluorescence lifetime (FLT) as an intrinsic discriminating parameter and resolving multi-exponential lifetime decays in flow cytometry. In our completed Phase I SBIR, we successfully demonstrated the automated removal of cellular AF interference in fluorophore- labeled human cells using time-resolved emission lifetimes using a 4-color system, establishing feasibility of the method. This Phase II grant will advance these findings towards commercialization by expanding the system into a fully operational 16-color Colorado analyzer with real-time data processing. The successful completion of the Colorado analyzer will lead to the first solution that automatically eliminates interference from AF in flow cytometer experiments. It will become a game-changing platform enabling cell biologists, immunologists, oncology researchers, diagnostic companies, and other FC users to obtain more accurate and quantitative results. Removal of cellular AF interference will yield enhanced sensitivity, which is important for detecting rare surface markers. This added capability of the Colorado platform will provide it with a significant competitive advantage over current commercial FC analyzers of similar multiplexing capabilities. This will enable the Colorado to replace current mid-level multiplexing instruments, which currently occupy a large swath of the rapidly growing $4.8 billion FC market.

项目摘要/摘要 流式细胞术是一种强大的细胞分析工具，它依赖于细胞的荧光标记。它可以提供 关于细胞计数、形状、大小、DNA含量、氧化还原状态、膜通透性和表面受体的信息， 在其他功能中。Fc是肿瘤生物学研究和临床研究中必不可少的工具 以及艾滋病诊断、干细胞治疗、免疫表型和癌症免疫治疗。领军人物之一 FC的局限性在于，外源添加的荧光团不是样品中唯一的荧光来源。 在某些情况下，自体荧光(AF)可能是每种情况下存在的信号的重要贡献者 探测通道经常与发射光谱重叠。房颤限制检测灵敏度及其原因 数据解释中的问题，这可能导致错误的科学结论。虽然有几种方法 已经被用来消除房颤，但没有令人满意的解决方案。 Kineic River一直在开发利用荧光寿命(FLT)作为本征的新方法 流式细胞术中参数识别和多指数寿命衰减的解析。在我们完成的 在第一阶段SBIR中，我们成功地展示了在荧光团中自动消除细胞房颤干扰- 使用四色系统使用时间分辨的发射寿命来标记人类细胞，建立了 方法。这项第二阶段的拨款将通过将该系统扩展到 完全可操作的16色科罗拉多分析仪，具有实时数据处理功能。成功地完成了 科罗拉多州分析仪将带来第一个自动消除流动中房颤干扰的解决方案 细胞仪实验。它将成为一个改变游戏规则的平台，使细胞生物学家、免疫学家、 肿瘤学研究人员、诊断公司和其他FC用户获得更准确和定量的 结果。消除细胞房颤干扰将产生更高的灵敏度，这对于检测罕见的 曲面标记。科罗拉多平台的这一新增功能将为其提供显著的竞争优势 比目前具有类似多路复用能力的商用FC分析仪更具优势。这将使 科罗拉多州将取代目前的中层多路复用仪器，后者目前占据了大片地区 快速增长的48亿美元的FC市场。