荧光寿命测定的现代方法与应用16.pptVIP

荧光寿命测定的现代方法与应用 七彩的荧光 荧光及荧光分光光度计的发展 荧光寿命的基本原理 荧光寿命测定的现代方法及数据处理 测定荧光寿命的一些应用 测定荧光寿命的仪器 主要内容 The emission of light from any substance is called luminescence. Some of the first scientific reports of luminescence appeared in the middle of the 18th century. In 1845, Herschel observed that an otherwise colourless solution of quinine (quinine absorbs in the UV region) in water emitted a blue colour under certain circumstances. In 1852 Sir G.G. Stokes studied the same compound and found that the emitted light has a longer wavelength than the light absorbed, the so-called Stokes’ shift. ( first described fluorescence ) In the 1920s and 1930s Jab?oński investigated polarized light and fluorescence and was able to show that the transition moments in absorption and emission are two different things. The Development of Fluorescence The first fluorescence microscopes were developed between 1911 and 1913 by German physicists Otto Heimstt and Heinrich Lehmann as a spin-off from the ultraviolet instrument. It wasnt until the early 1940s that Albert Coons developed a technique for labeling antibodies with fluorescent dyes, thus giving birth to the field of immunofluorescence. By the turn of the twenty-first century, the field of fluorescence microscopy was responsible for a revolution in cell biology, coupling the power of live cell imaging to highly specific multiple labeling of individual organelles and macromolecular complexes with synthetic and genetically encoded fluorescent probes. The Development of Fluorescence Microscopes Basic theory of fluorescence Lifetime The photophysical processes that occur from absorption to emission are often shown in a so-called Jab?oński diagram. 荧光是分子吸收能量后其基态电子S0被激发到单线激发态S1或S2后由第一单线激发态S1回到基态S0时所发生的。 荧光寿命是指分子在单线激发态 S1所平均停留的时间，或者说处于激发态S1的分子数目衰减到原来的1/e所经历的时间。 由于荧光现象多发生在纳秒级,这正好是分子运动所发生的时间尺度,因此利用荧光技术可以“看”到许多复杂的分子间作用过程,例如超分子体系中分子间的簇集、固液界面上吸附态高分子的构象重排、蛋白质高级结构的变化等。 荧光寿命的概念 荧光寿命测定的现代方法 荧光