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ISS ChronosBH Fluorescence Lifetime Spectrometer

ISS ChronosBH 荧光寿命光谱仪

基本信息

批准号：
7793245
负责人：
FRANCISCO J BEZANILLA
金额：
$ 21.23万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-02-18 至 2011-02-17
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Fluorescence spectroscopy has made myriad essential contributions to our understanding of molecular and cellular biology. Fundamental properties of intrinsic and extrinsic fluorophores probes in biological systems are exquisitely sensitive to the local bimolecular environment on (sub)-nanometer length scales. Measurement of fluorophore probe properties such as excited state fluorescence lifetime decay, polarization anisotropy, and energy transfer can reflect detailed structural, conformational, motional, binding, and dynamical interactions of macromolecules such as proteins, enzymes, RNA, and DNA. For instance, small changes in the fluorescence lifetime of a probe reveal the characteristics of processes in the local environment at short distances under 10 E (e.g., quenching and dielectric contributions) and long distances up to 100 E (e.g., Forster resonance energy transfer or FRET). This allows direct determination of important parameters of protein-protein binding, complex formation, molecular contact, proximity and overall conformational changes. These lifetime measurements require a highly sensitive lifetime fluorometer with picosecond resolution, such as the requested ISS ChronosBH Fluorescence Lifetime Spectrometer. This automated instrument combines the robust, fast, and proven Time-Correlated Single Photon Counting (TCSPC) method with a novel white light continuously tunable picosecond pulse excitation source that covers the entire visible to near-IR spectrum. The instrument can quickly measure multi-component lifetimes from 40ps to 1us (and longer in phosphorescence mode) with high fidelity from small volume (30uL) and weakly fluorescent samples. Kinetics studies are possible with a stop-flow apparatus. The instrument enables innovative new research as exemplified by 11 different projects from 9 major users. These projects encompass areas as diverse as molecular motions in ion channels, conformational changes in recombinase DNA complexes, amyloid proteins, catalysis regulation of human insulin degrading enzyme, and chemical synthesis of proteins for the development of new fluorescent probes. The need is urgent as there is no instrument of this capability at the University of Chicago or even the greater Chicago area. To increase availability and impact, the instrument will be placed in the interdisciplinary NanoBiology Facility shared core in the Institute for Biophysical Dynamics. Furthermore, it will be integrated in Graduate Biophysics lab curricula, thereby maximizing its exposure and impact on future biological and biomedical researchers.

描述(申请人提供)：荧光光谱学为我们理解分子和细胞生物学做出了许多重要贡献。生物体系中内在和外在荧光分子探针的基本性质对(亚)纳米尺度的局部双分子环境非常敏感。荧光探针性质的测量，如激发态荧光寿命衰减、偏振各向异性和能量转移，可以反映蛋白质、酶、RNA和DNA等大分子的详细结构、构象、运动、结合和动力学相互作用。例如，探测器荧光寿命的微小变化揭示了在10E以下的短距离(例如，猝灭和介电贡献)和高达100E的长距离(例如，Forster共振能量转移或FRET)的局部环境中的过程的特征。这使得可以直接确定蛋白质-蛋白质结合、复合体形成、分子接触、接近和整体构象变化的重要参数。这些寿命测量需要具有皮秒分辨率的高灵敏度寿命荧光仪，如所要求的ISS ChronosBH荧光寿命光谱仪。该自动化仪器结合了强大、快速且经过验证的时间相关单光子计数(TCSPC)方法和覆盖整个可见光至近红外光谱的新型白光连续可调皮秒脉冲激发源。该仪器可以从小体积(30uL)和弱荧光样品快速测量从40ps到1us的多组分寿命(在磷光模式下更长)，并具有高保真度。用停流装置进行动力学研究是可能的。该仪器能够进行创新的新研究，9个主要用户的11个不同项目就是例证。这些项目涵盖了离子通道中的分子运动、重组酶DNA复合体的构象变化、淀粉样蛋白、人胰岛素降解酶的催化调节以及用于开发新荧光探针的蛋白质的化学合成等领域。这种需求是迫切的，因为芝加哥大学甚至整个芝加哥地区都没有这种能力的工具。为了提高可用性和影响力，该仪器将放置在生物物理动力学研究所的跨学科纳米生物学设施共享核心中。此外，它将被整合到研究生生物物理学实验室课程中，从而最大限度地扩大其对未来生物和生物医学研究人员的接触和影响。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（3）

Ultrafast energy transfer from rigid, branched side-chains into a conjugated, alternating copolymer.

DOI：
10.1063/1.4855156
发表时间：
2014-01
期刊：
The Journal of chemical physics
影响因子：
0
作者：
G. Griffin;P. M. Lundin;B. Rolczynski;Alexander Linkin;Ryan D. McGillicuddy;Zhenan Bao;G. Engel
通讯作者：
G. Griffin;P. M. Lundin;B. Rolczynski;Alexander Linkin;Ryan D. McGillicuddy;Zhenan Bao;G. Engel

Cation-mediated optical resolution and anticancer activity of chiral polyoxometalates built from entirely achiral building blocks.