Time Resolved Fluorescence Spectroscopy

时间分辨荧光光谱是生物化学的一种强有力的工具，它可以为大分子络合物的结构和组装提供独特的见解。我们正在结束长达十年的研究，利用色氨酸(Trp)对水偶极运动引起的电场的皮秒响应来研究蛋白质附近的水的动力学。色氨酸是一种存在于大多数蛋白质中的氨基酸。TRP的上转换(子PS)研究的总体主题由我们在书中的章节中单独发表。所有的TRP研究现已转移到上海的东中国师范大学。 我们继续合作研究心肌线粒体的主要燃料-NADH的状态。我们的努力通过NADH的不同荧光寿命来区分NADH的游离群和结合群，并与光学显微镜核心公司和LCE合作，不断完善“衰变相关图像”软件，以更快地提取分离组织和/或肿瘤细胞内的NADH结合谱。今年，我们更新了用于大图像的DAI软件。我们继续为这个项目和其他项目开发时间分辨FCS中的耦合寿命和平移扩散能力。 我们使用我们的光谱工具帮助制作了一种用于细胞(组织)氧浓度检测的新成像分子的原型--我们将肌红蛋白修饰为毗邻红色荧光蛋白尾巴(例如mCherry)。它们通过改变肌红蛋白的寿命和亮度来响应肌红蛋白的氧结合。 我们还合作进行了超快研究，以帮助设计和开发荧光适配子，这是了解细胞RNA的新窗口。

Time-Resolved Fluorescence Spectroscopy is a powerful tool for biochemistry; it can provide unique insights into the structure and assembly of macromolecular complexes. We are wrapping up a decade of studying the dynamics of water near proteins using the picosecond response of Tryptophan(Trp), an amino acid found in most proteins, to the electric field caused by water dipole motions. The overall subject of upconversion (sub-ps) studies of Trp was separately published by us in book chapters. All Trp studies have now moved to East China Normal University in Shanghai. We continued collaborative studies into the status of a primary fuel of heart muscle mitochondria- NADH. Our efforts distinguish free and bound populations of NADH by their different fluorescence lifetimes, and in collaboration with Light Microscopy Core and LCE, we are continually refining 'Decay-Associated Images' software to more rapidly extract profiles of NADH binding within isolated tissue and/or tumor cells. This year we updated DAI software for large images. We continue to develop coupled lifetime and translational diffusion capabilities in time-resolved FCS for this and other projects. We used our spectroscopic tools to help prototype a new imaging molecule for cellular(tissue) oxygen concentration detection- we modified myoglobin to abut a red fluorescent protein tail (e.g. mCherry). They respond to the oxygen-binding of myoglobin by changing lifetime and brightness. We also collaborated on ultrafast studies to aid design and exploitation of fluorescent aptamers, a new window into cellular RNA.