Fluorometry

• Fluorometry procedure


  1. Requires a negative control (to set the zero point on the fluorometer)

  2. Requires a standard of known concentration

  3. The fluorometer shines light on the sample (excitation)

  4. The fluorometer measures level of fluorescent light being emitted to the side (at a 90º angle) of the excitation light beam

  
• Do not use glass (spectrophotometry) cuvettes in a fluorometer because the frosted glass on the side of the cuvette interferes with detection of fluorescent light
• Fluorometry

  Technique that measures fluorescence - light emitted by an atom/molecule after a finite duration subsequent to the absorption of electromagnetic energy
• Fluorescent dyes

  • Specifically bind to DNA/RNA
  • Fluorescence increases when bound to nucleic acid
• Fluorometry

  • Sensitive
  • Less susceptible to protein RNA contamination