- Perform Fluorescence and UV-Vis spectroscopy on the following second dye reaction solutions:
1. 70 Au/BSA ratio with dye
2. 166 Au/BSA ratio with dye
3. BSA/HCl with dye
- Prepared BSA/HCl control:
1. Mixed 1mL of HCl (2.84mM) + 1mL BSA(17.7µM) + 8mL water
2. Place into the oven (80 degrees) for 2 hours
- Preparation of dyed samples:
After dialysis, the following was observed:
The 70 ratio with dye was a clear solution with some purple bead like aggregates
The 166 ratio with dye was a clear solution with some purple fiber like aggregates
The BSA/HCl with dye was a pink solution with some pink bead shaped aggregates
1. Added Tris buffer until the solutions turned blue
2. Vortexed the samples: Aggregates in the 70 Au/BSA ratio and BSA/HCl solutions dissolved and remained in the 166 Au/BSA ratio solution
3. Centrifugation (5 minutes at 13200rpm) of the 166 ratio with dye solution to separate solution from aggregate
- UV-Vis Data - Second Dye Reaction
- Fluorescence Data - Second Dye Reaction
- When comparing this "second" dye reaction to the results obtained from the "first" dye reaction from 2/22/2012 there is consistency with respect to UV-Vis and inconsistency with respect to fluorescence.
- All UV-Vis spectra have the same trend in absorbance starting from the highest absorbance being just dye in water, followed by the BSA/HCl solution, then the 70 Au/BSA ratio solution, and finally the 166 Au/BSA ratio solution.
- During the 2/22 (First Dye) reaction, the highest emissions were the 70 Au/BSA ratio solution with dye and the BSA/HCl solution with dye. Both solutions emission peaks were very close to each other. The lowest emission was the 166 Au/BSA solution ratio with dye.
- During the 3/20 (Second Dye) reaction, the highest emission was the 166 Au/BSA ratio solution with dye. The lowest emissions were the 70 Au/BSA ratio solution with dye and the BSA/HCl solution with dye. These two emissions were very close to each other.
- Because the dye reaction is very sensitive to pH, it's possible that the pH is affecting the dye reaction and could be responsible for the inconsistencies. Further investigation into pH is necessary.