Objective
- Make stock concentrations (one set for both groups)
- Take UV-Vis and Fluorescence spectra for each stock
- Make a calibration curve for both fluorescence and UV-Vis
Procedure
Preparation of stock solutions
0.0120g of R6G in 10mL needed to make the initial stock solution of 2500uM, this was used to prepare a secondary stock of 500uM, used to make the dilutions for 0.1, 0.5, 0.75, 1.0, 1.2 and 1.5. The initial stock of 2500uM was used to make a 2uM. But the 1.5 and 2 uM were not used for the calibration curve because the fluorescence was too high that it did not provide accurate data.
- table of stock solution preparation
Concentration of stock solution (M)
|
Amount of Stock used (mL)
|
0.1 |
0.001
|
0.5 |
0.005
|
0.75 |
0.0075
|
1 |
0.01
|
1.2 |
0.012
|
Fluorescence
- Parameters of Fluorimeter:
- 500nm excitation
- 515-700nm scan range
- 10.0nm slit width
- 200nm/min scan speed
- Fluorescence spectra of standards
- Calibration curve based on Fluorescence
- Fluorescence of PVA/PVAC films and glass absorbance standards
UV-Vis
- UV-Vis spectra of standards
- Calibration curve based on UV-Vis
- UV-Vis of PVA/PVAC films and glass absorbance standards
|