Table 3.
TR-FRET reading settings using TECAN F500 plate reader
| Reading settings in Tecan F500 plate reader | |
|---|---|
| Fluorescence 1: d2 | |
| Excitation wavelength | 340 nm |
| Emission wavelength | 665 nm |
| Excitation bandwidth | 20 nm |
| Emission bandwidth | 8 nm |
| Gain | Optimal |
| Number of flashes | 10 |
| Integration time | 500 μs |
| Lag time | 150 μs |
| Fluorescence 2: Tb | |
| Excitation wavelength | 340 nm |
| Emission wavelength | 620 nm |
| Excitation bandwidth | 20 nm |
| Emission bandwidth | 10 nm |
| Gain | Optimal∗ |
| Number of flashes | 10 |
| Integration time | 500 μs |
| Lag time | 150 μs |
∗
When reading TR-FRET multiple times (every hour or in a kinetics mode, for example) the Gain of the donor signal should be set to a fixed value corresponding to the value obtained in the first reading.