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

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(Magnetic Stimulus Testing)
Current revision (15:26, 29 April 2014) (view source)
 
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==Objectives==
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{{deleteme}}
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#Magnetic testing/Fluorescence
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#Microscope (sonicated and oven samples prepared last week)
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#Prepare sonicated samples (Au incorporated)
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<br>
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==Microscope==
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*Please refer to Puja Mody's notebook for all microscope data
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<br>
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==Sample preparation==
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Table 1. FeSO<sub>4</sub> stock preparation
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Preparation of FeSO4 stock'''
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| align="center" style="background:#f0f0f0;"|''''''
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|-
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| Molecular Weight (g/mol)||278.01
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|-
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| mass(g)||0.83403
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|-
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| volume stock (L)||0.01
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|-
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| Concentration (M)||0.3
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|}
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<br>
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Table 2. BSA stock preparation
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Preparation of BSA stock'''
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| align="center" style="background:#f0f0f0;"|''''''
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|-
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| Molecular weight (g/mol)||66463
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|-
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| mass (g)||0.0174
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|-
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| volume stock (L)||0.01
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|-
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| Concentration (M)||2.618E-05
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|}
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<br>
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Table 3. Hemoglobin stock
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Preparation of hemoglobin Stock solution'''
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| align="center" style="background:#f0f0f0;"|''''''
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|-
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| Molecular weight (g/mol)||64500
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|-
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| mass hemglobin (g)||0.0995
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|-
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| volume solution (L)||0.1
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|-
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| Concentration (M)||1.54264E-05
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|}
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<br>
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Table 4. KNO<sub>3</sub> stock preparation
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Preparation of KNO3'''
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| align="center" style="background:#f0f0f0;"|''''''
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|-
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| Molecular Weight (g/mol)||101.1
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|-
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| mass(g)||0.9099
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|-
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| volume stock (L)||0.01
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|-
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| Concentration (M)||0.9
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|}
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<br>
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Table 5. KOH stock solution preparation
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Preparation of KOH stock'''
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| align="center" style="background:#f0f0f0;"|''''''
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|-
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| Molecular Weight (g/mol)||56.11
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|-
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| mass(g)||0.61721
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|-
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| volume stock (L)||0.01
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|-
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| Concentration (M)||1.1
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|}
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<br>
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Table 6. Gold stock solution preparation
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Preparation gold stock'''
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| align="center" style="background:#f0f0f0;"|''''''
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|-
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| Molecular weight Au (g/mol)||393.83
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|-
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| mass Au(g)||0.01446
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|-
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| volume stock(L)||0.01
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|-
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| Concentration (M)||0.003671635
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|}
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<br>
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Table 7.Magnetite:BSA preparation
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Ratio'''
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| align="center" style="background:#f0f0f0;"|'''Concentration BSA'''
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| align="center" style="background:#f0f0f0;"|'''Volume BSA Added (mL)'''
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| align="center" style="background:#f0f0f0;"|'''amount of FeSO4 added(mL)'''
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| align="center" style="background:#f0f0f0;"|'''amount of KOH added (mL)'''
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| align="center" style="background:#f0f0f0;"|'''amount of KNO3 added (mL)'''
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| align="center" style="background:#f0f0f0;"|'''volume Water added (mL)'''
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|-
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| 13000||2.30769E-06||0.881472149||1||0.909090909||4.444444444||2.764992498
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|}
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<br>
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Table 8.Magnetite:Hemoglobin preparation
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<br>
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Ratio'''
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| align="center" style="background:#f0f0f0;"|'''Concentration Hemoglobin'''
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| align="center" style="background:#f0f0f0;"|'''Volume Hemoglobin Added (mL)'''
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| align="center" style="background:#f0f0f0;"|'''amount of FeSO4 added(mL)'''
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| align="center" style="background:#f0f0f0;"|'''amount of KOH added (mL)'''
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| align="center" style="background:#f0f0f0;"|'''amount of KNO3 added (mL)'''
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| align="center" style="background:#f0f0f0;"|'''volume Water added (mL)'''
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|-
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| 16500||1.81818E-06||1.178620375||1||0.909090909||4.444444444||2.467844272
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|}
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<br>
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Table 9.Gold addition to Magnetite:BSA prep
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<br>
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Ratio'''
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| align="center" style="background:#f0f0f0;"|'''Concentration BSA'''
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| align="center" style="background:#f0f0f0;"|'''Volume BSA Added (mL)'''
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| align="center" style="background:#f0f0f0;"|'''Volume Gold (mL)'''
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| align="center" style="background:#f0f0f0;"|'''Volume Water(mL)'''
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|-
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| 130||1.92308E-06||0.734560124||0.680895574||8.584544302
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|}
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<br>
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Table 10.Gold addition to Magnetite:Hemoglobin prep
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<br>
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Ratio'''
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| align="center" style="background:#f0f0f0;"|'''Concentration Hemoglobin'''
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| align="center" style="background:#f0f0f0;"|'''Volume Hemoglobin Added (mL)'''
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| align="center" style="background:#f0f0f0;"|'''Volume Gold(mL)'''
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| align="center" style="background:#f0f0f0;"|'''Volume Water(mL)'''
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|-
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| 165||1.51515E-06||0.982183646||0.680895574||8.336920781
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|}
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<br>
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==Atomic Absorption==
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*Stock solutions were prepared for AA analysis tomorrow
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Table 1. Preparation of Iron standards
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<br>
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Concentration Standard (ug/mL)'''
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| align="center" style="background:#f0f0f0;"|'''Added Stock (mL)'''
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|-
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| 5||0.049850449
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|-
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| 10||0.099700897
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|-
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| 15||0.149551346
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|-
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| 20||0.199401795
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|-
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| 25||0.249252243
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|}
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<br>
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==Magnetic Stimulus Testing==
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'''General Protocol:'''
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#Centrifuge the 1.5mL eppendorf tubes with the samples for 15 minutes.
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#Remove R6G supernatant. Store in fresh eppendorf tube.
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#Add 1.3mL deionized H<sub>2</sub>O to the eppendorf tube.
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#Remove the deionized water.
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#Repeat this rinsing process a minimum of three times.
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#Add 1.3mL of water to the eppendorf tube.
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#Place sample on UV/fluorescence light to see if dye present in hydrogel samples
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#Centrifuge the sample for 15 minutes.
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#Remove the deionized water and place in clean eppendorf for testing (see if dye is coming out with centrifuging).
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#Add 1.5mL water to sample
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#Vortex the sample for ~2 min (until homogeneous suspension of hydrogels)
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#Apply magnet to outside of eppendorf tube for a minimum of 30 seconds (until all sample has aggregated).
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#Vortex sample again to re-suspend the hydrogels (minimum 15 seconds)
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#Re-apply magnet
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#Complete this vortex/magnet process for 4 minutes
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#Analyze deionized water using fluorescence
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<br>
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[[Image:Excitation and Emission Fluorescence spectra for magnetite-gold-protein hydrogels .png|500px]]
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<br>
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*All hydrogels responded to external magnetic stimuli. It appears sonication method was the most effective.
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<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
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