Difference between revisions of "DNA Melting: Model function and parameter estimation by nonlinear regression"
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* Only bound dye molecules may be excited. | * Only bound dye molecules may be excited. | ||
* An excited fluorophore will either bleach with probability <i>p</i> or return to the ground state with probability 1-<i>p</i>. | * An excited fluorophore will either bleach with probability <i>p</i> or return to the ground state with probability 1-<i>p</i>. | ||
| − | ** Mechanisms for returning to the ground state include fluorescence, phosphorescence, and non-radiative relaxation. | + | ** The constant 1-<i>p</i> includes all cases in which a molecule returns to the ground state unbleached. |
| + | ** Mechanisms for returning to the ground state include fluorescence, phosphorescence, and non-radiative relaxation. | ||
* Only excited fluorophore molecules bleach | * Only excited fluorophore molecules bleach | ||
* Bleaching is irreversible | * Bleaching is irreversible | ||
| + | * Magnitude of the illumination is equal to <i>I<sub>Ambient</sub></i> + <i>I<sub>LED</sub></i>, the intensities of ambient light and LED illumination, respectively. | ||
| + | |||
| + | ===Model=== | ||
| + | |||
| + | |||
{{Template:20.309 bottom}} | {{Template:20.309 bottom}} | ||
Revision as of 17:37, 13 November 2012
This subject matter will be explained in lecture, in tutorials, and in the lab.
Photobleaching model
Assumptions
- The dye is divided into two populations, bleached and unbleached, with concentrations S and S
- The initial dye concentration is S + S
- Bleached and unbleached molecules bind to dsDNA with the same affinity.
- Only bound dye molecules may be excited.
- An excited fluorophore will either bleach with probability p or return to the ground state with probability 1-p.
- The constant 1-p includes all cases in which a molecule returns to the ground state unbleached.
- Mechanisms for returning to the ground state include fluorescence, phosphorescence, and non-radiative relaxation.
- Only excited fluorophore molecules bleach
- Bleaching is irreversible
- Magnitude of the illumination is equal to IAmbient + ILED, the intensities of ambient light and LED illumination, respectively.
