HIGH-PRECISION FLOW CYTOMETRY OF BACTERIA AND VIRUSES

描述（由申请人提供）：大多数流式细胞仪均针对真核细胞分析进行了优化。传统仪器对细菌和病毒的测量受到光电子统计学的限制;因此，一些微生物成分和较小的微生物可能无法检测或无法精确测量。 在这个项目中，将建立和测试一个高灵敏度，高精度，慢流多参数流式细胞仪优化与微生物的工作。该设备将足够紧凑且用户友好，以放置在其可用于快速鉴定和表征多重抗性病原体和生物工程病原体以及开发用于这些病原体感染的治疗的环境中。将通过比较单光子计数和常规荧光检测方法，确定用于测量微生物核酸含量和细胞相关抗原或核酸探针的装置的灵敏度。 细菌pH值，膜电位和渗透性的比率测量的精度，如果可能的话，钙离子含量，将进行评估和比较，使用慢流和传统的仪器，以期完善细胞计数评估药物对细菌的影响。本文将探讨病毒染色强度与核酸类型和含量的关系，以期完善细胞计数法病毒检测和鉴定。

DESCRIPTION (provided by applicant): Most flow cytometers are optimized for analysis of eukaryotic cells. Measurements of bacteria and viruses in conventional instruments are limited by photoelectron statistics; some microbial constituents, and smaller microbes, may thus be undetectable or not measurable with precision. In this project, a high-sensitivity, high-precision, slow flow multiparameter flow cytometer optimized for work with microorganisms will be built and tested. The apparatus will be sufficiently compact and user-friendly to be placed in environments in which it can be used in rapid identification and characterization of multiply resistant and bioengineered pathogens and in development of therapy for infections with these pathogens. The sensitivity of the apparatus for measurement of microbial nucleic acid content and cell-associated antigens or nucleic acid probes will be established, comparing single photon counting and conventional fluorescence detection methods. The precision of ratiometric measurements of bacterial pH, membrane potential and permeability, and, if possible, calcium ion content, will be evaluated and compared using slow-flow and conventional instruments, with a view toward refining cytometric assessment of drug effects on bacteria. The relationship between staining intensity and nucleic acid type and content in viruses, will be investigated, with a view toward refining cytometric virus detection and identification.