User:Dhea Patel/Notebook/Hemoglobin Project/2014/02/05

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Setting Up Ocean Optics

  • Run Wizard for Absorbance Spectroscopy
  • Configure Data Collection as described below
  • Set Reference Blank and Dark
  • On the Absorbance view, click the "save" button to configure Save Settings
  • When you are ready to collect data, place the cuvette in the holder (with the cap) and click the "play" arrow followed by the "save" button.

OPD in H2O at 35C

  1. Stock solutions:
    1. In Lieu of Myoglobin
      1. 1.0mL H2O was kept in a separate centrifuge tube.
    2. OPD
      1. 3.8 mg OPD in 3.8mL H2O
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL H2O
  2. Experimental Run
    1. Sample chamber set to 35C and to stir
    2. 2.025mL OPD stock solution
    3. 0.75mL H2O2 stock solution
    4. Start Data Collection
      1. 1ms Integration
      2. 10 scan average
      3. Wait 1 minute after each scan
      4. Stop after 2 hours
      5. Files saved as: OPD_H2O2_H2O_35_SCANNUMBER.txt
    5. 0.225mL H2O added just before the third scan
      1. This is denoted in the data file for the third scan

OPD in H2O at 45°C

  1. The stock solutions were remade:
    1. 3.3 mg of OPD in 3.3 mL of H2O
    2. 0.114 mL of 30% H2O2 in 0.887mL H2O
  2. Experimental Run:
    1. Sample chamber set to 45°C and stir
    2. 2.025mL OPD stock solution
    3. 0.75mL H2O2 stock solution
    4. Start Data Collection
      1. 1ms Integration
      2. 10 scan average
      3. Wait 1 minute after each scan
      4. Stop after 2 hours
      5. Files saved as: OPD_H2O2_H2O_45_SCANNUMBER.txt
    5. 0.225mL H2O added just before the third scan
      1. This is denoted in the data file for the third scan