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Fluorescence Lifetimes

Pulse of Excited light fluorophore w/10ns decay fluorophore w/2ns decay. Fluorescence Lifetimes. k f. k i. 0. 400. 500. 600. 700. F. Short Wavelength Excitation. Long Wavelength Emission. Longer Wavelength. Higher Energy. INTENSITY. 400. 450. 500. 550. 600. 650.

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Fluorescence Lifetimes

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  1. Pulse of Excited light fluorophore w/10ns decay fluorophore w/2ns decay Fluorescence Lifetimes kf ki

  2. 0 400 500 600 700 F Short Wavelength Excitation Long Wavelength Emission Longer Wavelength Higher Energy INTENSITY 400 450 500 550 600 650 WAVELENGTH (nm) Fluorescence Fluorescence Emission Absorption Fluorescence Emission Absorption

  3. 0 400 500 600 700 INTENSITY 400 450 500 550 600 650 WAVELENGTH (nm) Fluorescence Emission is Sensitive to Solvent Conditions More polar conditions generally yield a RED SHIFT in emission Less polar conditions usually result in a BLUE SHIFT Increasing Solvent Polarity Longer Wavelength Higher Energy

  4. Solvent Effects

  5. Solvent Relaxation

  6. Solvent Relaxation of ANS

  7. Quinine Sulfate in 0%, 25%, 50% and 75% EtOH Intensity Change Wavelength Shift

  8. Lippert Equation

  9. Lifetime t0 = 1/G t= 1/(G + k) • Quantum Yield Q = t0/ t • Intensity Iem = Q·f(l)·Iex ·(l) ·C·Z

  10. Quantum Yield Measurements

  11. Inner Filter Effects

  12. Fluorescence Instrumentation

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