1 / 9

Fluorescence

Fluorescence. Energy diagram. Energy diagram. Quantum Yield - Q. Rate of depopulation. Red shift. Fluorimeter. Time resolved experiment . Spectrofluorimeter. Frequency domain (phase/modulation) . Excitation/ emmission spectrum :. Fluorophores. Intrinsic fluorophores. Extrinsic probes.

adonia
Download Presentation

Fluorescence

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Fluorescence Biophysical Tools '04 - Fluorescence

  2. Energy diagram Energy diagram Quantum Yield - Q Rate of depopulation Red shift Biophysical Tools '04 - Fluorescence

  3. Fluorimeter Time resolved experiment Spectrofluorimeter Frequency domain (phase/modulation) Excitation/emmission spectrum: Biophysical Tools '04 - Fluorescence

  4. Fluorophores Intrinsic fluorophores Extrinsic probes Solvent Effects Biophysical Tools '04 - Fluorescence

  5. Iװ I┴ Anisotropy Time resolved anisotropy Anisotropy Motional depolarization Biophysical Tools '04 - Fluorescence

  6. Phosphorescence Phosphorescence Øtriplet probes: eosin, erythrosin S T Biophysical Tools '04 - Fluorescence

  7. Anisotropic motion Hindered rotation Perrin Plot A(t) A∞ Biophysical Tools '04 - Fluorescence

  8. Quenching the rate of quenching = k[Q] Stern-Volmer plots 2 Fluor (no Q) / fluor (Q=x) 1 Quencher conc. Biophysical Tools '04 - Fluorescence

  9. 5 donor 4 donor/aceptor donor (AEDANS) 3 Intensity 2 acceptor 1 acceptor (AF) 0 500 700 400 600 Wavelength (nm) FRET Efficiency of transfer: E = rate of transfer /total rate of relaxation = E = kFET/(k + kFET ) = 1-tAD/tD = 1- FAD/FD dipole-dipole interactions kFET  (R-3)2 ~ 1/R6 Ro distance @ E=0.5 i.e. when kFET = 1/t then: E = R-6/(Ro-6 + R-6) Orientation problem: k2 Biophysical Tools '04 - Fluorescence

More Related