Fluorometry

• Photoluminescence 


  radiation of a longer wavelength than that originally absorbed is re-emitted after about 10^-8 seconds or longer
• Fluorescence emission 


  sample equally in all directions (90 degrees)
• Nephlometer 


  designed to measure the light scattered by a colloidal suspension
• MS Mass spectrometry 


  used to measure a wide range d clinically relevant analytes
• Mass spectrometer 


  ionizes a target molecule then separates and measures the mass of the molecule or its fragments
• m/z

  mass to charge
• Mass analysis

  process which a mixture of ionic species is identified according to m/z ratios
• Base peak 


  ion with highest abundance in the mass spectrum
• isotope dilution analysis 


  ability to quantify a compound relative to an isotopic species of known or fixed concentration
• IDMS 


  Isotope dilution mass spectrometry
• Mass spectrometer components

  • ion source
  • vacuum system
  • mass analyzer
  • detector
  • computer
• EI


  electronic ionization
• CI 


  chemical ionization
• HPLCMS 


  high performance liquid chromatograph mass spectrometer
• ESI


  electrospray ionization
• SSI

  Sonic spray ionization.
• APCI

  atmospheric pressure chemical ionization
• APPI

  atmospheric pressure photoionization
• Chemical Ionization


  reagent gas is ionized by electron beam
• Time of flight mass analyzer 


  measures the time it takes ions to move from ion source to the detector
• • QMF uses radio frequency (RF) - and direct current (DC) induced electric fields to separate ions.
• Matrix-Assisted Laser Desorption/lonization (MALDI)


  analyte is dissolved in a solution of matrix, which is a low molecular weight UV-absorbing compound.
• • QMF consists of four parallel rods in which the opposite pairs of rods are connected to the same DC and RF voltage.
• • TOF mass analyzer measures the time it takes ions of different masses to move from the ion source to the detector.
• • A lighter ion travels faster than a heavier ion, provided that both have the same kinetic energy.
• • A number of detectors are used to transform the intensity of the separated ions into an electronic signal.
• • The resulting signal is amplified 105 to 106.
• • Mass spectrometers coupled with gas and liquid chromatograph result in versatile analytical instruments that combine the resolving power of the chromatographs with the wonderful specificity and better sensitivity of a mass spectrometer.
• POS 


  point of sale
• Tagging 


  to make them more visible
• tagging 


  fluors fluorochrome
• fluorometer

  90 degrees perpendicular
• fluorometer parts

  • light source focusing lens
  • primary monochromator
  • sample cuvette
  • secondary monochromator
  • photo multiplier tube
  • signal voltage amplifier
  • display
  • reference beam photosensor
  • source supply
  • light block
• mercury 


  reference beam
• xenon 


  sample beam
• sources of light

  • reference beam
  • sample beam
• nephelometer 


  scattered light; measure light at right angle
• turbidimetry 


  turbid solution is blocking light
• dry slide technology 


  reflectance spectrophotometer