fluorescence polarization.docVIP

Fluorescence Spectroscopy ? Return to the Chemistry Home Page Fluorescence Schematic state energy level diagram S is singlet and T is triplet. The S0 state is the ground state and the subscript numbers identify individual states. Timescale S0 ? Sn absorption Sn ? S1 internal conversion (10-11 - 10-14 sec) S1 ? S0 + h? fluorescence (10-7 - 10-9 sec) S1 ? Tn intersystem crossing (10-8 sec) S1 ? S0 internal conversion (10-5 - 10-7 sec) T1 ? S0 + h? phosphorescence (10 - 10-3 sec) T1 ? S0 internal conversion (10 - 10-3 sec) ? Characteristics of Excited States Energy Lifetime Quantum Yield Polarization ? Phosphorescence occurs at longer wavelength than does fluorescence Often, the emission band is red-shifted relative to the absorption band: Stokes shift Excited states decay exponentially with time I = I0e-t/? I0 is the initial intensity at time zero, I is the intensity at some later time t, and ? is the lifetime of the excited state. Also, kF = 1/??, where kF is the rate constant for fluorescence. ? Quantum Yield = ? ?F = number of fluorescence quanta emitted divided by number of quanta absorbed to a singlet excited state ?F = ratio of photons emitted to photons absorbed Quantum yield is the ratio of photons emitted to photons absorbed by the system: ?F = kF / kF + kISC + knr + kq + kr ? Polarization Molecule of interest is randomly oriented in a rigid matrix (organic solvent at low temperature or room temperature polymer). Plane polarized light is used as the excitation source. The degree of polarization is defined as follows: where I|| and I? are the intensities of the observed parallel and perpendicular components, respectively. ? is the angle between the emission and absorption transition moments. If ? is 0° than P = +1/2, and if ? is 90° than P = -1/3. Figure 26, Becker, pg. 84 Polarization of fluorescence of phenol in propylene glycol at -70°C shows that the transition moments of the corresponding absorption bands are mutually perpendicular. P