Fluorimeter for biophysical characterizations

点击翻译按钮获取中文摘要

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.