User:Matthew R Skorski/Notebook/471 - Exp BioChem/2016/01/27

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Objective

Today's objective is to practice synthesizing AuNP+Rhodamine fibers. We will also practice adding Rhodamine at different concentrations and at different points in the synthesis.

Protocol

First, we used volumetric flasks to make the following stock solutions of gold, lysozyme, and Rhodamine in DI water:

  • 50mL of 3316.µM gold stock solution
    • Molecular weight of gold solid is 339.79g/mol
  • 50mL of 38.30µM lysozyme stock solution
    • Molecular weight of lysozyme solid is 14307g/mol
  • 25mL of 1079µM Rhodamine stock solution


Our goal was to make 36 1mL fiber samples. For each sample:

  • The concentration of gold would be 250µM (i.e. we'd add 75.39µL of our gold stock)
  • The concentration of lysozyme would be 5.556µM (i.e. we'd add 145.07µL of our lysozyme stock)
  • We would bring the total volume up to 1mL using DI water (i.e. we'd add 779.54µL to each sample)

These gold and lysozyme concentrations should be ideal for forming fibers.


We were also going to add Rhodamine to the fibers at different points in the synthesis in order to determine which time allowed the maximum percentage of Rhodamine to be incorporated into the fibers. Additionally, were were going to add varying concentrations of Rhodamine to determine the best concentration to use for maximum incorporation of the dye into the fibers.

  • We did three serial dilutions of the 1079µM stock of Rhodamine (diluted by a factor of 10 for each dilution). This way, we could change the concentration of Rhodamine added to each sample but keep the volume of Rhodamine added constant.
    • The volume of Rhodamine added to each sample would be 9.27µL
  • NOTE: we assumed the volume of Rhodamine added to each sample would be negligible, so when doing concentration calculations, we assumed the total volume of each sample would be 1000µL (even though technically it would be 1000µL-9.27µL).


We made the following samples to test these two variables:


Sample # Time in Synthesis that Rhodamine is Added Concentration of Rhodamine
1 Before heating samples in oven 0µM (control)
2 Before heating samples in oven 0µM (control)
3 Before heating samples in oven 0µM (control)
4 Before heating samples in oven 0.01µM
5 Before heating samples in oven 0.01µM
6 Before heating samples in oven 0.01µM
7 Before heating samples in oven 0.1µM
8 Before heating samples in oven 0.1µM
9 Before heating samples in oven 0.1µM
10 Before heating samples in oven 1µM
11 Before heating samples in oven 1µM
12 Before heating samples in oven 1µM
13 When samples get cloudy (partway though heating) 0µM (control)
14 When samples get cloudy (partway though heating) 0µM (control)
15 When samples get cloudy (partway though heating) 0µM (control)
16 When samples get cloudy (partway though heating) 0.01µM
17 When samples get cloudy (partway though heating) 0.01µM
18 When samples get cloudy (partway though heating) 0.01µM
19 When samples get cloudy (partway though heating) 0.1µM
20 When samples get cloudy (partway though heating) 0.1µM
21 When samples get cloudy (partway though heating) 0.1µM
22 When samples get cloudy (partway though heating) 1µM
23 When samples get cloudy (partway though heating) 1µM
24 When samples get cloudy (partway though heating) 1µM
25 After samples have cooled 0µM (control)
26 After samples have cooled 0µM (control)
27 After samples have cooled 0µM (control)
28 After samples have cooled 0.01µM
29 After samples have cooled 0.01µM
30 After samples have cooled 0.01µM
31 After samples have cooled 0.1µM
32 After samples have cooled 0.1µM
33 After samples have cooled 0.1µM
34 After samples have cooled 1µM
35 After samples have cooled 1µM
36 After samples have cooled 1µM


For the control samples, we were going to add 9.27µL of DI water instead of Rhodamine.


Because of time, we ended up not adding Rhodamine to samples 13-24. Matt and Michael added Rhodamine to samples 25-36 on Thursday.

Future Plans

In the future, our goal will be to synthesize the fibers on Tuesdays and to analyze the fibers and supernatants on Wednesdays. We will come in early on Tuesdays to start the synthesis so that we have enough time to add Rhodamine to samples 13-36.