Difference between revisions of "User:Moira M. Esson/Notebook/CHEM-571/2014/04/15"

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(Magnetic Stimulus Testing)
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==Objectives==
+
{{deleteme}}
#Magnetic testing/Fluorescence
 
#Microscope (sonicated and oven samples prepared last week)
 
#Prepare sonicated samples (Au incorporated)
 
<br>
 
==Microscope==
 
*Please refer to Puja Mody's notebook for all microscope data
 
<br>
 
==Sample preparation==
 
Table 1. FeSO<sub>4</sub> stock preparation
 
{| {{table}}
 
| align="center" style="background:#f0f0f0;"|'''Preparation of FeSO4 stock'''
 
| align="center" style="background:#f0f0f0;"|''''''
 
|-
 
| Molecular Weight (g/mol)||278.01
 
|-
 
| mass(g)||0.83403
 
|-
 
| volume stock (L)||0.01
 
|-
 
| Concentration (M)||0.3
 
|}
 
<br>
 
Table 2. BSA stock preparation
 
{| {{table}}
 
| align="center" style="background:#f0f0f0;"|'''Preparation of BSA stock'''
 
| align="center" style="background:#f0f0f0;"|''''''
 
|-
 
| Molecular weight (g/mol)||66463
 
|-
 
| mass (g)||0.0174
 
|-
 
| volume stock (L)||0.01
 
|-
 
| Concentration (M)||2.618E-05
 
|}
 
<br>
 
Table 3. Hemoglobin stock
 
{| {{table}}
 
| align="center" style="background:#f0f0f0;"|'''Preparation of hemoglobin Stock solution'''
 
| align="center" style="background:#f0f0f0;"|''''''
 
|-
 
| Molecular weight (g/mol)||64500
 
|-
 
| mass hemglobin (g)||0.0995
 
|-
 
| volume solution (L)||0.1
 
|-
 
| Concentration (M)||1.54264E-05
 
|}
 
<br>
 
Table 4. KNO<sub>3</sub> stock preparation
 
{| {{table}}
 
| align="center" style="background:#f0f0f0;"|'''Preparation of KNO3'''
 
| align="center" style="background:#f0f0f0;"|''''''
 
|-
 
| Molecular Weight (g/mol)||101.1
 
|-
 
| mass(g)||0.9099
 
|-
 
| volume stock (L)||0.01
 
|-
 
| Concentration (M)||0.9
 
|}
 
<br>
 
Table 5. KOH stock solution preparation
 
{| {{table}}
 
| align="center" style="background:#f0f0f0;"|'''Preparation of KOH stock'''
 
| align="center" style="background:#f0f0f0;"|''''''
 
|-
 
| Molecular Weight (g/mol)||56.11
 
|-
 
| mass(g)||0.61721
 
|-
 
| volume stock (L)||0.01
 
|-
 
| Concentration (M)||1.1
 
|}
 
<br>
 
Table 6. Gold stock solution preparation
 
{| {{table}}
 
| align="center" style="background:#f0f0f0;"|'''Preparation gold stock'''
 
| align="center" style="background:#f0f0f0;"|''''''
 
|-
 
| Molecular weight Au (g/mol)||393.83
 
|-
 
| mass Au(g)||0.01446
 
|-
 
| volume stock(L)||0.01
 
|-
 
| Concentration (M)||0.003671635
 
|}
 
<br>
 
Table 7.Magnetite:BSA preparation
 
{| {{table}}
 
| align="center" style="background:#f0f0f0;"|'''Ratio'''
 
| align="center" style="background:#f0f0f0;"|'''Concentration BSA'''
 
| align="center" style="background:#f0f0f0;"|'''Volume BSA Added (mL)'''
 
| align="center" style="background:#f0f0f0;"|'''amount of FeSO4 added(mL)'''
 
| align="center" style="background:#f0f0f0;"|'''amount of KOH added (mL)'''
 
| align="center" style="background:#f0f0f0;"|'''amount of KNO3 added (mL)'''
 
| align="center" style="background:#f0f0f0;"|'''volume Water added (mL)'''
 
|-
 
| 13000||2.30769E-06||0.881472149||1||0.909090909||4.444444444||2.764992498
 
|}
 
<br>
 
Table 8.Magnetite:Hemoglobin preparation
 
<br>
 
{| {{table}}
 
| align="center" style="background:#f0f0f0;"|'''Ratio'''
 
| align="center" style="background:#f0f0f0;"|'''Concentration Hemoglobin'''
 
| align="center" style="background:#f0f0f0;"|'''Volume Hemoglobin Added (mL)'''
 
| align="center" style="background:#f0f0f0;"|'''amount of FeSO4 added(mL)'''
 
| align="center" style="background:#f0f0f0;"|'''amount of KOH added (mL)'''
 
| align="center" style="background:#f0f0f0;"|'''amount of KNO3 added (mL)'''
 
| align="center" style="background:#f0f0f0;"|'''volume Water added (mL)'''
 
|-
 
| 16500||1.81818E-06||1.178620375||1||0.909090909||4.444444444||2.467844272
 
|}
 
<br>
 
Table 9.Gold addition to Magnetite:BSA prep
 
<br>
 
{| {{table}}
 
| align="center" style="background:#f0f0f0;"|'''Ratio'''
 
| align="center" style="background:#f0f0f0;"|'''Concentration BSA'''
 
| align="center" style="background:#f0f0f0;"|'''Volume BSA Added (mL)'''
 
| align="center" style="background:#f0f0f0;"|'''Volume Gold (mL)'''
 
| align="center" style="background:#f0f0f0;"|'''Volume Water(mL)'''
 
|-
 
| 130||1.92308E-06||0.734560124||0.680895574||8.584544302
 
|}
 
<br>
 
Table 10.Gold addition to Magnetite:Hemoglobin prep
 
<br>
 
{| {{table}}
 
| align="center" style="background:#f0f0f0;"|'''Ratio'''
 
| align="center" style="background:#f0f0f0;"|'''Concentration Hemoglobin'''
 
| align="center" style="background:#f0f0f0;"|'''Volume Hemoglobin Added (mL)'''
 
| align="center" style="background:#f0f0f0;"|'''Volume Gold(mL)'''
 
| align="center" style="background:#f0f0f0;"|'''Volume Water(mL)'''
 
|-
 
| 165||1.51515E-06||0.982183646||0.680895574||8.336920781
 
|}
 
<br>
 
==Atomic Absorption==
 
*Stock solutions were prepared for AA analysis tomorrow
 
Table 1. Preparation of Iron standards
 
<br>
 
{| {{table}}
 
| align="center" style="background:#f0f0f0;"|'''Concentration Standard (ug/mL)'''
 
| align="center" style="background:#f0f0f0;"|'''Added Stock (mL)'''
 
|-
 
| 5||0.049850449
 
|-
 
| 10||0.099700897
 
|-
 
| 15||0.149551346
 
|-
 
| 20||0.199401795
 
|-
 
| 25||0.249252243
 
|}
 
<br>
 
 
 
==Magnetic Stimulus Testing==
 
'''General Protocol:'''
 
#Centrifuge the 1.5mL eppendorf tubes with the samples for 15 minutes.
 
#Remove R6G supernatant. Store in fresh eppendorf tube.
 
#Add 1.3mL deionized H<sub>2</sub>O to the eppendorf tube.
 
#Remove the deionized water.
 
#Repeat this rinsing process a minimum of three times.
 
#Add 1.3mL of water to the eppendorf tube.
 
#Place sample on UV/fluorescence light to see if dye present in hydrogel samples
 
#Centrifuge the sample for 15 minutes.
 
#Remove the deionized water and place in clean eppendorf for testing (see if dye is coming out with centrifuging).
 
#Add 1.5mL water to sample
 
#Vortex the sample for ~2 min (until homogeneous suspension of hydrogels)
 
#Apply magnet to outside of eppendorf tube for a minimum of 30 seconds (until all sample has aggregated).
 
#Vortex sample again to re-suspend the hydrogels (minimum 15 seconds)
 
#Re-apply magnet
 
#Complete this vortex/magnet process for 4 minutes
 
#Analyze deionized water using fluorescence
 
<br>
 
[[Image:Excitation and Emission Fluorescence spectra for magnetite-gold-protein hydrogels .png|500px]]
 
<br>
 
*All hydrogels responded to external magnetic stimuli. It appears sonication method was the most effective.
 
 
 
 
 
 
 
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|}
 
|}
  
 
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