荧光寿命测定现代方法与应用.pptVIP

的荧光 主要内容 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 The oment of Fluorescence 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 GG. 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 Jablonski investigated polarized light and fluorescence and was able to show that the transition moments in absorption and emission are two different things The Dev ol Fluorescence Microscopes 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 technique for labeling antibodies with fluorescent dves. thus giving birth to the field of immunofluorescence By the turn of the twenty-first century, the field of uorescence microscopy was responsible Tor a revolution In coupling the power of live cell imaging to highly specific multiple labeling of individual organelles and acromolecular complexes with synthetic and genetically scent Basic theory of fluorescence Lifetime The photophysical processes that occur from absorption to emission are often shown in a so-called Jablonski diagram ablonski Energy Diagram Excitation (Absorption) Excited singlet States 10Seconds Vibrational Energy States Internal Internal Conversion onversion and vibrational Delayed Relaxation Fluorescence 1 10-10sec) 5 Excited 2 Triplet 1 state Fluorescence 10--10sec) Intersystem Crossing intersystem Crossing Non-Radiative Relaxation [Triplet) Quenching Phosphorescence Non-Radiative so 【10--10-