User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/07/25: Difference between revisions
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==July 25, 2013== | ==July 25, 2013== | ||
===Procedure Update: MHA Silica-Coating=== | |||
According to [http://link.springer.com/content/pdf/10.1007%2Fs11051-012-0758-z.pdf Aggregation and adhesion of gold nanoparticles | |||
in phosphate buffered saline], PBS and prolonged sonication actually cause aggregation in gold nanoparticle solutions: "results showed an optimized sonication time of 10 min to reach equilibrium for plain gold nanoparticles in water." Instead of using PBS to dilute all components, filtered MilliQ water will be used to dilute nanoparticles as well as MHA, as the organic solvent in solution prior to the actual silica-coating may have also been affecting observed the gold nanoparticle aggregation. | |||
*Originally detailed on [http://openwetware.org/wiki/User:Madeleine_Y._Bee/Notebook/Single_Molecule_Fluorescence/2013/05/24 05/24/2013] | |||
*Last updated on [http://openwetware.org/wiki/User:Madeleine_Y._Bee/Notebook/Single_Molecule_Fluorescence/2013/07/12#Procedure_Update:_Silica-coating_AuNPs_by_MHA 07/12/2013] | |||
*Day One: | |||
**Attach MHA to AuNPs: add 5mL of 10mM MHA to 5mL of AuNPs, stir overnight | |||
***To prepare 10mM MHA solution: | |||
***(5 mL)×(L/1000mL)×(10mmoles/L)×(1mol/1000mmoles)×(288.49g/mol)=0.0144g MHA in 5mL filtered MilliQ water | |||
***To prepare 5mL of 9.4nM AuNPs: | |||
***(29.67nM)V=(9.4nM)(5mL) | |||
***V=1.584mL AuNPs in 3.416mL filtered MilliQ water | |||
***Combine MHA and AuNP solutions | |||
*Day Two: | |||
**Prepare the following solutions: | |||
#1mL 9.4nM, 10nm AuNP&MHA/20μL TES/0.125mL NH4OH/5mL propanol | |||
#1mL 9.4nM, 10nm AuNP&MHA/15μL TES/0.125mL NH4OH/5mL propanol | |||
#1mL 9.4nM, 10nm AuNP&MHA/10μL TES/0.125mL NH4OH/5mL propanol | |||
#1mL 9.4nM, 10nm AuNP&MHA/5μL TES/0.125mL NH4OH/5mL propanol | |||
#*Stir solutions for one hour to allow reaction to proceed | |||
#*Centrifuge solutions at 4000 rpm for one hour | |||
#*Re-suspend particles in 140uL PBS (per solution) | |||
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Revision as of 07:07, 25 July 2013
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July 25, 2013Procedure Update: MHA Silica-CoatingAccording to [http://link.springer.com/content/pdf/10.1007%2Fs11051-012-0758-z.pdf Aggregation and adhesion of gold nanoparticles in phosphate buffered saline], PBS and prolonged sonication actually cause aggregation in gold nanoparticle solutions: "results showed an optimized sonication time of 10 min to reach equilibrium for plain gold nanoparticles in water." Instead of using PBS to dilute all components, filtered MilliQ water will be used to dilute nanoparticles as well as MHA, as the organic solvent in solution prior to the actual silica-coating may have also been affecting observed the gold nanoparticle aggregation.
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