NOVEL FLUORESCENCE METHODS FOR BIOMEDICAL APPLICATIONS

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。我们的研究小组正在应用先进的荧光方法来研究相关的生物医学过程。通过与一家专门从事荧光技术的工业合作伙伴的合作，我们可以获得新型激光诱导荧光仪器，该仪器可以捕获波长时间矩阵（WTMs），该矩阵由一系列发射波长产生的荧光衰减曲线组成。我们快速收集和分析wtm的能力激发了对寿命对环境敏感的染料的研究。这样的吗?一生探针吗?我们的工业合作伙伴开发的价格合理的仪器为简化探针设计和扩展SNP检测和定量PCR的多路复用策略提供了机会。项目前9个月的大部分时间用于创建开展研究所需的基础设施，以及培训本科生操作生物材料、数据收集和数据分析。研究实验室的面积扩大了一倍，并进行了全面翻新。基本的实验室项目（即冷冻室，高压灭菌器，多通道移液器）和一个定制修改的商业荧光计与终身使用能力交付并安装。学院对分子生物学家的寻找工作已顺利结束，该人员自2005年1月起已完全融入研究项目。四名本科生在本学年每季度参与一次研究；其中三名学生将于2005年夏季进行全日制研究。三名新生（一名大三学生和两名大二学生）计划在秋季加入该项目。对激光染料的寿命进行了初步的研究。寿命测量精度达到10皮秒以上。对更具挑战性的终身探测目标系统的初步研究正在进行中。在接下来的一年里，我们计划建立一个寿命探针库，并改进评估染料作为寿命探针候选物的程序。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Our research group is applying advanced fluorescence methodology to study relevant biomedical processes. Via collaboration with an industrial partner that specializes in fluorescence technology, we have access to novel laser induced fluorescence instrumentation that can capture wavelength-time matrices (WTMs), which consist of fluorescence decay curves generated at a series of emission wavelengths. Our ability to rapidly collect and analyze WTMs motivates the search for dyes whose lifetimes are environmentally sensitive. Such ?lifetime probes? and affordable instrumentation developed by our industrial partner opens up opportunities to simplify probe design and expand multiplexing strategies for SNP detection and quantitative PCR. Much of the first nine months of the project was taken up with creating the infrastructure necessary to conduct the research and with training the undergraduate students in manipulation of biological materials, data collection, and data analysis. The research laboratory was doubled in size and completely renovated. Essential laboratory items (i.e. freezer, autoclave, multi-channel pipets) and a custom modified commercial fluorimeter with lifetime capability were delivered and installed. A faculty search for a molecular biologist was successfully concluded and this person has been fully integrated into the research project since January 2005. Four undergraduates have been participating in the research on a quarter-time basis during the academic year; three of these students will conduct full-time research in the summer of 2005. Three new students (one junior and two sophomores) plan to join the program in the fall. Preliminary lifetime studies have been conducted on laser dyes. The desired lifetime measurement precision of ten picoseconds or better has been met. Initial studies into the more challenging lifetime probe-target systems are underway. During the next year we plan to develop a library of lifetime probes and improve procedures to evaluate dyes as candidates for lifetime probes.