IMPROVEMENTS IN FLOW CYTOMETRY OF BACTERIA & VIRUSES

细菌流式细胞术的改进

• 批准号: 3498487
• 负责人: HOWARD M. SHAPIRO
• 金额: $ 4.84万
• 依托单位: HOWARD M. SHAPIRO, MD, P.C.
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-03-01 至 1990-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3498487
• 关键词: biomedical equipment resource; clinical biomedical equipment; fluorescent dye /probe; light scattering; microorganism classification

Several approaches to increasing the sensitivity of flow cytometry for detection and characterization of small bacteria and large virus particles will be explored, including a water-immersed flow chamber, a laser Doppler light scatter detector system, front end electronics incorporating source noise compensation, and light noise reduction by acousto-optic modulation. A high-sensitivity flow cytometer developed as a result of these studies will be used for measurements of light scattering and fluorescence form bacteria, viruses, and test particles smaller than 0.5 mum in diameter. Two-parameter measurements of light scattering and cyanine dye fluorescence on multiple aliquots of a sample will be used to estimate bacterial membrane potentials and to rapidly determine which of several energy sources would support the growth of an organism and which of several inhibitors would prevent growth. Ratiometric measurements of the fluorescence of the fluorescein derivative BCECF will be used to detect pH gradients between bacterial cytoplasm and the medium. Multistation, multiparameter measurements of light scattering and of the fluorescence of mixtures of dyes, including membrane potential and pH probes, fluorescent stains for protein, DNA and RNA, and fluorogenic enzyme substrates, will be investigated to determine which parameters add the most additional information relevant to bacterial identification.

提高流式细胞术检测肿瘤细胞的灵敏度的几种方法 小细菌和大病毒颗粒的检测和表征 将探索，包括水浸流动室，激光多普勒 光散射检测器系统，前端电子设备结合源 噪声补偿和通过声光调制降低光噪声。 作为这些研究的结果， 将用于光散射和荧光形式的测量 细菌、病毒和直径小于0.5 μ m的测试颗粒。 光散射和菁染料荧光的双参数测量 将使用多份样品的多个等分试样来估计细菌 膜电位，并迅速确定几种能量 一个生物体的生长需要几种来源， 抑制剂会阻止生长。 的比率测量 荧光素衍生物BCECF的荧光将用于检测pH 细菌细胞质和培养基之间的梯度。 多站， 光散射和荧光的多参数测量 染料的混合物，包括膜电位和pH探针，荧光 蛋白质、DNA和RNA以及荧光酶底物的染色剂将被 调查，以确定哪些参数添加最多的额外 与细菌鉴定相关的信息。