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Лекция 3

Лекция 3. Влияние внутри и межмолекулярных взаимодействий на люминесценцию. Растворители: универсальные взаимодействия. реактивное. макроскопическое. Специфические взаимодействия. Неоднородное уширение спектров. Динамическое и статическое тушение люминесценции. Ассоциация. ПЭЭВ.

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Лекция 3

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  1. Лекция 3

  2. Влияние внутри и межмолекулярных взаимодействий на люминесценцию

  3. Растворители: универсальные взаимодействия

  4. реактивное макроскопическое

  5. Специфические взаимодействия

  6. Неоднородное уширение спектров

  7. Динамическое и статическоетушение люминесценции

  8. Ассоциация

  9. ПЭЭВ

  10. ISS PC1 (ISS Inc., Champaign, IL, USA) Fluorolog-3 (Jobin Yvon Inc, Edison, NJ, USA ) QuantaMaster (OBB Sales, London, Ontario N6E 2S8)

  11. Fluorometer Components Excitation Polarizer Sample Emission Polarizer Excitation Wavelength Selection Light Source Emission Wavelength Selection Computer Detector Fluorometer: The Basics Note: Both polarizers can be removed from the optical beam path

  12. The Laboratory Fluorometer Pex Pem Pem Standard Light Source: Xenon Arc Lamp Exit Slit ISS (Champaign, IL, USA) PC1 Fluorometer

  13. Light Sources

  14. Xenon Arc Lamp Profiles Lamp Light Sources Ozone Free • Xenon Arc Lamp (wide range of wavelengths) • High Pressure Mercury Lamps (High Intensities but concentrated in specific lines) • Mercury-Xenon Arc Lamp (greater intensities in the UV) • Tungsten-Halogen Lamps • Light emitting diodes (LEDs) Multiple color LEDs can be bunched to provide a broad emission range) Visible UV Mercury-Xenon Arc Lamp Profile

  15. Light Emitting Diodes (LED) Wavelengths from 350 nm to 1300 nm Near UV LED

  16. 528nm 532nm 543nm 514nm 633nm 442nm 325nm 488nm 351 nm 364 nm 295nm 576nm Lasers Light Sources Titanium:Sapphire 690 nm – 990 nm 200 300 400 500 600 700 Wavelength (nm) Green He-Ne 10 mW Orange He-Ne 10 mW Argon-ion 100 mW Helium-cadmium Nd-YAG He-Ne >10 mW

  17. Laser Diodes

  18. Wavelength Selection Fixed Optical Filters Tunable Optical Filters Monochromators

  19. Hoya O54 Long Pass Optical Filters Transmission (%) Spectral Shape Thickness Physical Shape Fluorescence (!?) Wavelength (nm)

  20. Tunable Optical Filters Liquid Crystal Filters: An electrically controlled liquid crystal elements to select a specific visible wavelength of light for transmission through the filter at the exclusion of all others. AO Tunable Filters: The AOTF range of acousto-optic devices are solid state optical filters. The wavelength of the diffracted light is selected according to the frequency of the RF drive signal. Isomet (http://www.isomet.com/index.html)

  21. Monochromators Mirrors Czerny-Turnerdesign • Slit Width (mm) is the dimension of the slits. • Bandpass is the FWHM of the selected wavelength. • The dispersion is the factor to convert slit width to bandpass. Exit Slit Entrance slit Rotating Diffraction Grating (Planar or Concaved)

  22. The Inside of a Monochromator Mirrors Grating Nth Order (spectral distribution) Zero Order (acts like a mirror)

  23. Higher Order Light Diffraction Emission Scan: Excitation 300 nm Glycogen in PBS 2nd Order Scatter (600 nm) Excitation (Rayleigh) Scatter (300 nm) Fluorescence (au) 2nd Order RAMAN (668 nm) Water RAMAN (334 nm) Wavelength (nm) Fluorescent Contaminants

  24. Detectors Scallop Scallop Eyes From http://www.eyedesignbook.com/index.html Image courtesy of BioMEDIA ASSOCIATES http://www.ebiomedia.com

  25. The Classic PMT Design Vacuum Dynodes Photocathode e- e- l e- e- e- e- Anode e- e- e- e- e- e- e- Window Constant Voltage (use of a Zenor Diode) Current Output resister series (voltage divider) Ground capacitor series (current source) High Voltage Supply (-1000 to -2000 V)

  26. Hamamatsu R928 PMT Family R2949 Window with Photocathode Beneath

  27. PMT Quantum Efficiencies Cathode Material Window Material

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