Table 2.
Criterion | Description | Quantification |
---|---|---|
A | fluorescence fluctuations must be fast enough | λ ≥ 0.1 s−1 |
B | diffusion through the evanescent wave must be fast enough | λ ≤ 500 s−1 |
C | magnitude of Gs(τ) is high enough | Gs(0) ≥ 0.05 |
D | Gs(τ) is not overwhelmed by Ga(τ) | Gs(0)/Ga(0) ≥ 0.1 |
E | surface rebinding is negligible | P ≤ 0.05 |
This table describes the criteria used to determine conditions for which TIR-FCS can measure thermodynamic and kinetic parameters describing the interaction of fluorescent molecules with surface binding sites given that the mechanism is a simple bimolecular reaction as defined in Eq. 2.