Scanning Microfluorescence Analysis of Immobilized-Cell Biocatalysts

这项提案的目的是开发和优化一种新的 实验方法，基于扫描显微荧光， 将细胞生理学表征为时间和 并定位在固定化细胞生物催化剂内。 的 然后将使用一种方法研究质粒稳定性 在固定化大肠杆菌细胞中。 该方法采用 生物催化剂配置，其允许目视观察 在离营养源不同距离处捕获的细胞， 这样营养良好的细胞和饥饿的细胞 查看 最近开发的扫描显微荧光 仪器也在使用。 该仪器具有先进的图像- 分析能力，其激发激光束可以是 聚焦到直径小于一微米。 这些特征 允许单个细菌细胞的荧光被 测定了 质粒稳定性将采用 荧光标记基因连接到靶质粒上。 这 新的分析方法既强大又通用。 一 各种各样的固定化细胞现象可以通过 更换荧光染色剂。 研究结果将 加强对固定化细胞的基本认识 生理学，并允许在数学模型的改进， 工业上重要的固定化细胞方法。

The object of this proposal is to develop and optimize a novel experimental approach, based on scanning microfluorescence, to characterize cellular physiology as a function of both time and position within immobilized-cell biocatalysts. The approach will then be used to investigate plasmid stability within immobilized Escherichia coli cells. The method employs a biocatalyst configuration which allows visual observation of cells entrapped at various distances from the nutrient supply, so that well-nourished, as well as starving cells may be viewed. A recently developed scanning microfluorescence instrument is also used. This instrument has advanced image- analysis capabilities, and its excitation laser beam can be focused to a diameter less than one micron. These features allow fluorescence of individual bacterial cells to be measured. Plasmid stability will be monitored using fluorescent marker genes attached to target plasmids. This new analytical approach is both powerful and versatile. A wide variety of immobilized-cell phenomena can be studied by changing fluorescent stains. Results of the study will enhance the fundamental understanding of immobilized-cell physiology and allow refinements in mathematical models for industrially important immobilized-cell processes.