Влияние внутри и межмолекулярных взаимодействий на люминесценцию

Влияние внутри и межмолекулярных взаимодействий на люминесценцию

Растворители: универсальные взаимодействия

макроскопическое реактивное

Специфические взаимодействия

Неоднородное уширение спектров

Динамическое и статическое тушение люминесценции

Ассоциация

ПЭЭВ

ISS PC 1 (ISS Inc. , Champaign, IL, USA) Fluorolog-3 (Jobin Yvon Inc, Edison, NJ, USA ) Quanta. Master (OBB Sales, London, Ontario N 6 E 2 S 8)

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

The Laboratory Fluorometer Standard Light Source: Xenon Arc Lamp Exit Slit Pex Pem ISS (Champaign, IL, USA) PC 1 Fluorometer

Light Sources

Xenon Arc Lamp Profiles Lamp Light Sources Ozone Free 1. Xenon Arc Lamp (wide range of wavelengths) 2. High Pressure Mercury Lamps (High Intensities but concentrated in specific lines) UV Visible 3. Mercury-Xenon Arc Lamp (greater intensities in the UV) 4. Tungsten-Halogen Lamps 5. Light emitting diodes (LEDs) Multiple color LEDs can be bunched to provide a broad emission range) Mercury-Xenon Arc Lamp Profile

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

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

Laser Diodes

Wavelength Selection Fixed Optical Filters Tunable Optical Filters Monochromators

Transmission (%) Long Pass Optical Filters Spectral Shape Thickness Physical Shape Fluorescence (!? ) Wavelength (nm) Hoya O 54

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)

Monochromators Mirrors Czerny-Turner design 1. Slit Width (mm) is the dimension of the slits. 2. Bandpass is the FWHM of the selected wavelength. Exit Slit 3. The dispersion is the factor to convert slit width to bandpass. Entrance slit Rotating Diffraction Grating (Planar or Concaved)

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

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

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

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

Hamamatsu R 928 PMT Family R 2949 Window with Photocathode Beneath

PMT Quantum Efficiencies Cathode Material Window Material