User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/06/26: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Reading and FCS Data</span> | ||
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===Notes from Literature=== | ===Notes from Literature=== | ||
Looking for information on silica-coating thickness for DNA attachment procedure. | Looking for information on silica-coating thickness for DNA attachment procedure. | ||
*From: http://pubs.acs.org/doi/pdf/10.1021/nn401775e | *From: [http://pubs.acs.org/doi/pdf/10.1021/nn401775e Distance and Wavelength Dependent Quenching of Molecular Fluorescence by Au@SiO2 Core-Shell Nanoparticles] | ||
**References 8-13, 15, 26 | **References 8-13, 15, 26 | ||
**"Gold nanoparticles between 10 and 20 nm in diameter are important because they are too small to scatter light significantly | **"Gold nanoparticles between 10 and 20 nm in diameter are important because they are too small to scatter light significantly[...]fluorescence of molecules in close proximity to these particles is quenched." | ||
**"...almost complete (>90%) quenching at short separations to negligible electromagnetic coupling at a distance of 43 nm" | **"...almost complete (>90%) quenching at short separations to negligible electromagnetic coupling at a distance of 43 nm" | ||
**"...hydrolysis of the reactive NHS moiety on the dye is much slower in ethanol than in water." (For attachment procedure) | **"...hydrolysis of the reactive NHS moiety on the dye is much slower in ethanol than in water." (For attachment procedure) | ||
**"...attachment was more efficient for particles with a thicker silica shell due to the higher surface area and the increased number of binding sites available per particle." | **"...attachment was more efficient for particles with a thicker silica shell due to the higher surface area and the increased number of binding sites available per particle." | ||
**"For average dye separations above 15 nm (less than 30 dyes per particle) | **"For average dye separations above 15 nm (less than 30 dyes per particle)[...]small (<5%) decrease in fluorescence lifetime and in a 20% decrease in steady-state fluorescence relative to the fluorescence of the free dye. A very pronounced decrease in both fluorescence intensity and lifetime is observed for dye separations below 10 nm (more than 75 dyes per particle). For the highest dye loading, the steady-state fluorescence decreases by more than 90% and the fluorescence lifetime is reduced to 0.5 ns compared to 4.0 ns for the free dye in solution." | ||
*Solvents: ethanol for coated spheres, 2-propanol for uncoated | *Solvents: ethanol for coated spheres, 2-propanol for uncoated | ||
===FCS Measurements=== | ===FCS Measurements=== | ||
*Samples prepared: | *Samples prepared to be added to data and calibration graphs from 06/25/2013: | ||
**(1) 100pM DNA/120pM MB | **(1) 100pM DNA/120pM MB | ||
**(2) 80pM DNA/96pM MB | **(2) 80pM DNA/96pM MB | ||
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**(7) 80pM Oligo D | **(7) 80pM Oligo D | ||
**(8) 60pM Oligo D | **(8) 60pM Oligo D | ||
*Data added to calibration curves from 06/25/2013, but samples will be made and run again due to inconsistencies | |||
===DNA/ThT Sample Prep=== | |||
*Another 700nM sample was made to complete the calibration curve for the DNA/ThT samples | |||
*Fluorescence measurements taken | |||
*Results still inconclusive, will be redone tomorrow | |||
Revision as of 13:15, 26 June 2013
Reading and FCS Data | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
June 26, 2013Notes from LiteratureLooking for information on silica-coating thickness for DNA attachment procedure.
FCS Measurements
DNA/ThT Sample Prep
|