User:Moira M. Esson/Notebook/CHEM-571/2014/04/15

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Revision as of 11:24, 29 April 2014 by Moira M. Esson (talk | contribs) (Magnetic Stimulus Testing)
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Objectives

  1. Magnetic testing/Fluorescence
  2. Microscope (sonicated and oven samples prepared last week)
  3. Prepare sonicated samples (Au incorporated)


Microscope

  • Please refer to Puja Mody's notebook for all microscope data


Sample preparation

Table 1. FeSO4 stock preparation

Preparation of FeSO4 stock '
Molecular Weight (g/mol) 278.01
mass(g) 0.83403
volume stock (L) 0.01
Concentration (M) 0.3


Table 2. BSA stock preparation

Preparation of BSA stock '
Molecular weight (g/mol) 66463
mass (g) 0.0174
volume stock (L) 0.01
Concentration (M) 2.618E-05


Table 3. Hemoglobin stock

Preparation of hemoglobin Stock solution '
Molecular weight (g/mol) 64500
mass hemglobin (g) 0.0995
volume solution (L) 0.1
Concentration (M) 1.54264E-05


Table 4. KNO3 stock preparation

Preparation of KNO3 '
Molecular Weight (g/mol) 101.1
mass(g) 0.9099
volume stock (L) 0.01
Concentration (M) 0.9


Table 5. KOH stock solution preparation

Preparation of KOH stock '
Molecular Weight (g/mol) 56.11
mass(g) 0.61721
volume stock (L) 0.01
Concentration (M) 1.1


Table 6. Gold stock solution preparation

Preparation gold stock '
Molecular weight Au (g/mol) 393.83
mass Au(g) 0.01446
volume stock(L) 0.01
Concentration (M) 0.003671635


Table 7.Magnetite:BSA preparation

Ratio Concentration BSA Volume BSA Added (mL) amount of FeSO4 added(mL) amount of KOH added (mL) amount of KNO3 added (mL) volume Water added (mL)
13000 2.30769E-06 0.881472149 1 0.909090909 4.444444444 2.764992498


Table 8.Magnetite:Hemoglobin preparation

Ratio Concentration Hemoglobin Volume Hemoglobin Added (mL) amount of FeSO4 added(mL) amount of KOH added (mL) amount of KNO3 added (mL) volume Water added (mL)
16500 1.81818E-06 1.178620375 1 0.909090909 4.444444444 2.467844272


Table 9.Gold addition to Magnetite:BSA prep

Ratio Concentration BSA Volume BSA Added (mL) Volume Gold (mL) Volume Water(mL)
130 1.92308E-06 0.734560124 0.680895574 8.584544302


Table 10.Gold addition to Magnetite:Hemoglobin prep

Ratio Concentration Hemoglobin Volume Hemoglobin Added (mL) Volume Gold(mL) Volume Water(mL)
165 1.51515E-06 0.982183646 0.680895574 8.336920781


Atomic Absorption

  • Stock solutions were prepared for AA analysis tomorrow

Table 1. Preparation of Iron standards

Concentration Standard (ug/mL) Added Stock (mL)
5 0.049850449
10 0.099700897
15 0.149551346
20 0.199401795
25 0.249252243


Magnetic Stimulus Testing

General Protocol:

  1. Centrifuge the 1.5mL eppendorf tubes with the samples for 15 minutes.
  2. Remove R6G supernatant. Store in fresh eppendorf tube.
  3. Add 1.3mL deionized H2O to the eppendorf tube.
  4. Remove the deionized water.
  5. Repeat this rinsing process a minimum of three times.
  6. Add 1.3mL of water to the eppendorf tube.
  7. Place sample on UV/fluorescence light to see if dye present in hydrogel samples
  8. Centrifuge the sample for 15 minutes.
  9. Remove the deionized water and place in clean eppendorf for testing (see if dye is coming out with centrifuging).
  10. Add 1.5mL water to sample
  11. Vortex the sample for ~2 min (until homogeneous suspension of hydrogels)
  12. Apply magnet to outside of eppendorf tube for a minimum of 30 seconds (until all sample has aggregated).
  13. Vortex sample again to re-suspend the hydrogels (minimum 15 seconds)
  14. Re-apply magnet
  15. Complete this vortex/magnet process for 4 minutes
  16. Analyze deionized water using fluorescence


Excitation and Emission Fluorescence spectra for magnetite-gold-protein hydrogels .png

  • All hydrogels responded to external magnetic stimuli. It appears sonication method was the most effective.