Development of a Dual-Beam Multi-Parameter Flow Cytometer for Microbial Ecology

这里描述的仪器开发工作将大大 扩大和增强我们现有的能力（“原型”） 高灵敏度流式细胞仪分析天然 微生物种群 具体而言，我们建议： .安装氩氪混合气体激光器， 激发波长的选择。 .添加双光束所需的光学器件和电子器件 激发和4通道检测。 .开发一个高度灵活的基于 现成的电子模块 .将流式细胞仪与基于实时神经网络的 数据分析系统 该工具将用于制定识别和 自然样品中特定微生物的计数（基于 16S rRNA靶向的寡核苷酸探针），并在 原位生理参数（使用更传统的 细胞化学技术）。 还将使用流式细胞仪 支持正在进行的细菌细胞周期调控研究 和微藻。

The instrument development work described here will greatly expand and enhance the capabilities of our existing ("prototype") high sensitivity flow cytometer for the analysis of natural microbial populations. Specifically, we propose to: . Install an argon-krypton mixed gas laser to provide a wide choice of excitation wavelengths. . Add the optics and electronics required for dual beam excitation and 4 channel detection. . Develop a highly flexible data acquisition system based on off-the-shelf electronics modules. . Interface the flow cytometer with a real-time neural net-based data analysis system. The instrument will be used to develop methods of identifying and enumerating specific microorganisms in natural samples (based on 16S rRNA-targeted oligonucleotide probes), and of analyzing in situ physiological parameters (using more traditional cytochemical techniques). The flow cytometer will also be used to support ongoing studies of cell cycle regulation in bacteria and microalgae.