Fluorimeter for biophysical characterizations

荧光光谱（荧光光谱）是一种技术，我们使用它来更好地了解生物大分子（蛋白质，DNA，RNA）的构象如何决定功能。通过测量荧光团（一种在给定波长的光激发下发出荧光的分子）发出的光，我们可以获得有关蛋白质内部或表面局部化学环境的有用信息。荧光团可以是固有的（例如，作为蛋白质一部分的氨基酸）或外源性（例如，附着在蛋白质上的荧光染料）。我们使用荧光测定法来报告蛋白质相关荧光团被掩埋或淬灭的程度，因为从这样的荧光团发出的光的量可以由于蛋白质构象变化而急剧增加或减少。使用称为荧光分光光度计（荧光计）的仪器进行荧光测量。我们需要一个荧光计来表征我们蛋白质的结构性质，并了解它们如何相互作用。在过去的一年里，我们部门唯一的研究专用荧光计在经过11年的多次维修后完全停止了运作。我们迫切需要一种具有偏振和温度控制能力的现代荧光计来推进我们的研究计划。荧光偏振的使用是我们荧光分析需要的核心，因为它具有探测蛋白质构象变化和蛋白质-蛋白质相互作用的能力。此外，我们中的许多人需要精确的温度控制，以了解温度变化如何影响我们的蛋白质或蛋白质-荧光团缀合物的荧光特性。

Fluorescence spectroscopy (flourimetry) is a technique that we use to better understand how the conformation of biological macromolecules (protein, DNA, RNA) can determine function. By measuring light emitted from a flourophore (a molecule that fluoresces upon excitation by light of a given wavelength) we can obtain useful information on the local chemical environments either within or on the surface of a protein. A fluorophore can either be intrinsic (e.g., an amino acid that is part of a protein) or extrinsic (e.g., a fluorescent dye attached to a protein). We use fluorimetry to report on the extent to which a protein-associated fluorophore is buried or quenched, since the amount of light emitted from such a fluorophore can increase or decrease dramatically as a result of protein conformational changes. Fluorescence measurements are performed with an instrument called a fluorescence spectrophotometer (fluorimeter). We need a fluorimeter to characterize the structural nature of our proteins as well as to gain an understanding of how they interact with each other. In the past year, our Department's single research-dedicated fluorimeter completely ceased functioning after 11 years and many repairs. We urgently require a modern fluorimeter with polarization and temperature control capabilities to advance our research programs. The use of fluorescence polarization is central to our fluorimetry needs given its power in probing protein conformational changes and protein-protein interactions. Also, many of us require accurate temperature control in order to see how changes in temperature affect fluorescence properties of our proteins or protein-fluorophore conjugates.