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After you assembled the fibrils, don't rush to let the QD labeled streptavidin sit on the steats. If there are a huge amount of fibrils, even though the number of seats on each fibril is sufficiently large, there may not be enough QD's to fully occupy them and hence result in a low average intensity on each fibril. On the contary, if we first dilute the buffer of fibrils, QD's may be able to fully occupy each of the fibril and hence result in a high average intensity on each fibril.
After you assembled the fibrils, don't rush to let the QD labeled streptavidin sit on the seats. If there are a huge amount of fibrils, even though the number of seats on each fibril is sufficiently large, there may not be enough QD's to fully occupy them and hence result in a low average intensity on each fibril. On the contary, if we first dilute the buffer of fibrils, QD's may be able to fully occupy each of the fibrils and hence result in a high average intensity on each fibril.
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Revision as of 10:14, 18 October 2011

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 <img src="http://openwetware.org/images/5/53/NBgamers_team_logo.png" alt="NBgamers (Team of NanoBiotechnology)" title="NBgamers (Team of NanoBiotechnology)">
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<h3>Intensity Optimization</h3>

<p> In our project, each individual fibril is labeled by quantum dots (QD). The intensity of the output fluorescence signal from the QD's determines the ability of indentifying the fibrils. Therefore, maximizing the QD fluorescence signal intensity is one of our tasks. There are several factors influencing the output intensity: </p>

<p> 1. The ratio between the free Aβ monomers and biotinylated Aβ monomers. </p>

<p> Imagining you put the free Aβ monomers and the biotinylated Aβ monomers into buffers to let them spontaneously assemble into fibrils. A large proportion of free ones may result in a relatively long fibril length distribution, but since QD labeled streptavidin only "sits" on biotinylated Aβ monomers, the average intensity on each fibril may be low. </p>

<p> 2. The dilution factor of fibrils. </p>

<p> After you assembled the fibrils, don't rush to let the QD labeled streptavidin sit on the seats. If there are a huge amount of fibrils, even though the number of seats on each fibril is sufficiently large, there may not be enough QD's to fully occupy them and hence result in a low average intensity on each fibril. On the contary, if we first dilute the buffer of fibrils, QD's may be able to fully occupy each of the fibrils and hence result in a high average intensity on each fibril. </p>

<p> The bar chart below shows the measured output intensity from the QD's under different conditions: </p>

<p align="center"> <a href="http://openwetware.org/images/6/6a/Intensity_Optimization.png"> <img src="http://openwetware.org/images/6/6a/Intensity_Optimization.png" width="454.5" height="373.5" alt="Intensity measurement (Clikc to enlarge.)" title="Intensity measurement (Clikc to enlarge.)"> </a> </p>

<p> The result of the intensity optimization gives us a suggestion: A condition with 20% biotin ratio and 50X dilution factor should be a good choice. </p> <br />

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