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INTRODUCTION TO FLUORESCENCE (two-photon, three steps, phase incoherent process)

INTRODUCTION TO FLUORESCENCE (two-photon, three steps, phase incoherent process). Einstein’s coefficients, dipole transitions. Experimental setup T-configuration Jablonsky diagram Franck-Condon principle, reflection rule and Stokes shift Lifetime and quantum yield

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INTRODUCTION TO FLUORESCENCE (two-photon, three steps, phase incoherent process)

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  1. INTRODUCTION TO FLUORESCENCE (two-photon, three steps, phase incoherent process) • Einstein’s coefficients, dipole transitions. • Experimental setup • T-configuration • Jablonsky diagram • Franck-Condon principle, reflection rule and Stokes shift • Lifetime and quantum yield • Fluorescence quenching (collisional) • London’s theory of molecular forces • RET resonant Energy Transfer • FRAP (Fluorescence Recovery after Photo Bleaching

  2. Experimental setup

  3. Gregorio Weber’s T-configuration

  4. Stokes shift

  5. Protein Intrinsic Fluorophores

  6. stati di tripletto e di singoletto

  7. Kasha’s rule: emission spectra are independent of the excitation wavelength.

  8. Quantum yield Q and lifetime τ Γ=spectral emissivity, knr rate of non radiative transitions teoricamente

  9. Fluorescence Quenching Collisional quenching Stern-Volmer equation

  10. Fluorescence Anisotropy anisotropy polarization

  11. What is lost in steady state measurements?

  12. Green-fluorescent proteins GFP

  13. Non radiative energy transfer (RET)

  14. Center for fluorescence spectroscopy (J. Lakowicz) http://cfs.umbi.umd.edu/ ISS http://www.iss.com/ Fluorescence foundation http://www.fluorescence-foundation.org/ Enrico Gratton’s homepage http://www.lfd.uci.edu/~gratton/ Laboratory of fluorescence dynamics Gregorio Weber http://www.lfd.uci.edu/weber/ CORSO DI MICROSCOPIA OTTICA DI CLEGG http://online.physics.uiuc.edu/courses/phys598om/spring12/

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