TABLE 3.
Lifetimes and FRET efficiencies estimated by means of FLIM for internal complexes
Fluorescence was monitored from 19 cells in which the localization of eGFP-M2 and eYFP-M2 was predominantly internal. The traces from binned pixels were analyzed in terms of Equations 22 and 23 as described in the legend to Table 2. Distributions of lifetimes from two exponentials (Equation 22, q = 2) required two gaussians for 4 cells and three gaussians for 19 cells (Equation 23, n = 2 or 3). Efficiencies were calculated from τDA and τD for each cell (Equation 24), and the individual values were averaged to obtain the means (±S.E.) listed in the table.
| q (Equation 22) | q (Equation 23) | Lifetime |
Efficiency (Equation 24) | |||
|---|---|---|---|---|---|---|
| τ0,1 (τDA) | τ0,2 (τD) | τ0,3 (τA) | ||||
| ns | ns | ns | ||||
| A | 2 | 2 (19) | 2.15 ± 0.08 (19) | 2.78 ± 0.09 (19) | 0.23 ± 0.04 | |
| B | 2 | 2 (4) | 2.02 ± 0.07 (19) | 2.44 ± 0.03 (19) | 2.98 ± 0.10 (15) | 0.17 ± 0.03 |
| 3 (15) | ||||||
| C | 3 | 3 (19) | 2.08 ± 0.09 (19) | 2.55 ± 0.09 (19) | 3.17 ± 0.12 (15) | 0.18 ± 0.05 |