NOVEL FLUORESCENCE METHODS FOR BIOMEDICAL APPLICATIONS

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心机构，不一定为研究者机构。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合作，我们获得了新型激光诱导荧光仪器，该仪器可以捕获波长-时间矩阵（WTM），其中包括在一系列发射波长下产生的荧光衰减曲线。我们快速收集和分析WTM的能力促使我们寻找寿命对环境敏感的染料。这样的？寿命探测器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。 该项目前九个月的大部分时间都用于创建进行研究所需的基础设施，并培训本科生操作生物材料，数据收集和数据分析。研究实验室的规模扩大了一倍，并进行了全面翻新。交付并安装了基本的实验室物品（即冷冻箱、高压灭菌器、多通道移液管）和一台定制的改进型商用荧光计（具有终身使用能力）。一个教师寻找分子生物学家成功地结束了，这个人已经完全融入研究项目自2005年1月以来。四名本科生在本学年期间每四分之一时间参加研究;其中三名学生将在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四学生）计划在秋季加入该项目。 对激光染料进行了初步的寿命研究。所需的寿命测量精度为10皮秒或更好已经得到满足。目前正在对更具挑战性的寿命探针-靶系统进行初步研究。在明年，我们计划开发一个图书馆的寿命探针和改进程序，以评估染料作为候选人的寿命探针。

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.