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| Dual color FCS measurements contain a wealth of information about molecular interactions and stoichiometry. In general, 5 peices of information are measured in a typical experiment: green intensity, red intensity, green brightness (or G(0)), red brightness (or G(0)), and dual color brightness (or cross correlation G(0)). As a result, 5 parameters can be measured in the experiment. Below, we have implemented javascript calculators for FCS and N&B analysis given two different scenarios. In both scenarios, we have no red to green bleedthrough and the green to red bleedthrough is defined as the ratio of the green species intensity in the red channel to the green species intensity in the red channel. In both scenarios, we have three species present: an independent green species, an independent red species, and an interacting species. In the first scenario we know the stoichiometry of the bound complex and we calculate the number of all three species in the focal volume as well as the brightness of the green and red independent species. In the second scenario, we know the brightnesses of the green and red independent species and we calculate the number of all three species in the focal volume as well as the stoichiometry of the interacting complex. |
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Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
FCS
PCH
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Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
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Download Presentation |
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Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
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Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
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Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
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The calculators are built using html forms and javascript. If you look at the source code for this page, you should be able to find java script functions like jcalc...(). The functions used for these calculations are listed below.
Note that ng (the stoichiometry of the green species) is dependent on nr. In the same way, εgm (brightness of the green monomer) is dependent on εrm. β is the green to red bleedthrough fraction mentioned above. For calculations involving G(0), the amplitudes are converted first to brightnesses (see the Brightness Analysis page) and then the calculation proceeds with the equations above..
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Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
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