User:Madeleine Y. Bee/Notebook/CHEM-581 Experimental Chemistry/2014/10/01

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October 1, 2014


R6G Fluorescence: Calibration and Measurement

  1. Make stock concentrations (both groups can use the same solutions)
    • Initial stock: 2500uM, secondary stock: 500uM
    • 0.10uM
      • Made with 500uM solution
    • 0.50uM
      • Made with 500uM solution
    • 1.0uM
      • Made with 500uM solution
    • 1.5uM
      • Made with 500uM solution
    • 2.0uM
      • Made with 2500uM solution
    • 0.75uM
      • Made with 500uM solution
    • 1.2uM
      • Made with 500uM solution
  2. Take UV-Vis and Fluorescence spectra of these samples
    • Fluorimeter Settings:
      • 500nm excitation
      • 515-700nm scan range
      • 10.0nm slit width
      • 200nm/min scan speed
  3. Make a calibration curve based on UV-Vis.
    • Compare your data to some published values
  4. Make a calibration curve based on the fluorescence.
    • In order to do this, you'll need to measure the area under the fluorescence curve, not just the fluorescence peak height.


Calibration Curve: R6G Absorbance

2014 1001 R6G calibration.PNG

Measurement Results: R6G Absorbance

2014 1001 R6G abs.PNG

Calibration Curve: R6G Fluorescence

2014 1003 R6G solutions fluor calibration.PNG

Measurement Results: R6G Fluoresence

2014 1001 R6G fluor.PNG