User:Patrick Hampson/Notebook/chem471/2016/09/07

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Fluorescence 201697

Protocol

  1. Starting the Peltier controller
    1. Make sure that the ribbon cord connects the peltier controller and the cuvette holder
    2. Fill the chiller bucket with water
    3. Place the aquarium pump (with tubes attached to the Peltier heat exchanger) into the bucket and plug in the pump so that water flows
    4. Turn on the peltier controller (switch in the back of the instrument)
    5. Use the arrow buttons to set the temperature.
  2. Turning off the Peltier controller
    1. Turn the controller power off
    2. Unplug the aquarium pump
    3. Dump the water from the bucket into the sink


Sample Preparations

stock solutions

  1. BSA
    1. 25.33 mg of myoglobin with water in 10.0 mL volumetric flask
    2. MW = 66776 g/mol
    3. concentration = 37.93 μM

samples

  1. BSA Fluorescence
    1. 3 mL total
      1. 0.25 mM Au
      2. 3.125 μM BSA
      3. HCl or NaOH appropriate for your pH
      4. water
  2. BSA for UV-Vis during next session
    1. 5 mL total
      1. 0.25 mM Au
      2. 3.125 μM BSA
      3. HCl or NaOH appropriate for your pH
      4. Appropriate amount of fructose
      5. water


Fluorescence data

Fluorescence summary 160909.PNG This graph was comprised from fluorescent scans taken once every five minutes. There is a steady increase in the peaks of the data over the course of time.


Emission maximum 160909.PNG After the initial datapoint, there is a drastic increase in emissions. This increase is not constant, as the rest of the data collected throughout the duration of the experiment were relatively similar. The datapoints appeared to form a plateau, however upon closer inspection, there is a slight increase in the emissions as time goes on.


Integrated Fluorescence.PNG

This graph displays the integrated fluorescence over time. The data resembles a minor parabola for the first 50 minutes of the experiment. The data then flattens out to just below 5,000 until it reaches 100 minutes. At 100 minutes there is a severe spike in the data, and integrated fluorescence increases to over 20,000. There is a steady linear increase in the integrated fluorescence for the remainder of the experiment.

UV-Vis

  1. 5 mL total
    1. 0.25 mM Au
    2. 3.125 μM BSA
    3. HCl or NaOH appropriate for your pH
    4. Appropriate amount of fructose
    5. water


stock solutions

  1. The NaOH and HCl samples were made exactly as designed.
  2. AuCl3
    1. mass = 38.5 mg
    2. 25.0 mL volumetric flask
    3. concentration = 5.08 mM
  3. Fructose
    1. mass = 23.48 mg
    2. 10.0 mL volumetric flask
    3. concentration = 13.0 mM
  4. Bovine Serum Albumin
    1. mass = 25.33 mg
    2. 10.0 mL volumetric flask
    3. concentration = 37.93 mM
0.125 mM Fructose
pH μL of 1 mM HClstock μL of 1 mM NaOHstock μL of 1 M NaOHstock μL of AuCl3stock μL of Fructosestock μL of BSAstock μL of Water
4 500 0 0 246 48 408 3798
5 50 0 0 246 48 408 4248
6 5 0 0 246 48 408 4293
7 0 0 0 246 48 408 4298
8 0 5 0 246 48 408 4293
9 0 50 0 246 48 408 4248
10 0 500 0 246 48 408 3798
11 0 0 5 246 48 408 4293
12 0 0 50 246 48 408 4248
0.25 mM Fructose
pH μL of 1 mM HClstock μL of 1 mM NaOHstock μL of 1 M NaOHstock μL of AuCl3stock μL of Fructosestock μL of BSAstock μL of Water
4 500 0 0 246 96 408 3750
5 50 0 0 246 96 408 4200
6 5 0 0 246 96 408 4245
7 0 0 0 246 96 408 4250
8 0 5 0 246 96 408 4245
9 0 50 0 246 96 408 4200
10 0 500 0 246 96 408 3570
11 0 0 5 246 96 408 4245
12 0 0 50 246 96 408 4200


Solutions before being heated in the oven (low reactivity): 160907 stock solutions 671 lab.jpg

  • Matt Hartings 14:02, 9 September 2016 (EDT): I remade your samples because you got strange results. Here's my description