Molecular Fluorescence Instrumental : Part 3: Standard Curve

At two excitation wavelengths (both ~370 and ~440 nm) you determine the exact wavelengths to use.

1. Double click the Simple Reads icon on the desktop.
2. Click on the Setup button, and set the emission wavelength and excitation wavelength in the window.
3. Set the excitation and emission slits at 10 nm, and the Average Time at 5 seconds.
4. Click on Read to begin data acquisition.
5. Change the excitation wavelength and repeat the procedure.
6. Do this for all your standards, and repeat three times for the unknown sample.
7. Print the data files when you are done.
8. When you are done with the fluorimeter, exit the programs, switch out the instrument, and remember to sign the logbook.
9. Create a standard curve in Excel or Google sheets to determine the concentration of Riboflavin in your unknown solution.