Table 1.
Confocal Imaging Settings for QD–Dye FRET Pairs
| QD–Dye | Excitation, nm | Emission, nm | Images: Emission Filters |
|---|---|---|---|
| QD566–AF568 | Acceptor (543) | LP590 (590–700) | A |
| Donor (488) | BP535-590 (535–590) | qD | |
| Donor (488) | LP590 (590–700) | uFRET | |
| QD–Dye | Excitation, nm | Emission, nm | Images: Linear Unmixinga |
| QD580–AF594 | Acceptor (514) | λ stack (505–719) | A |
| Donor (458) | λ stack (505–719) | qD | |
| uFRET |
a
Images were generated by linear unmixing to separate acceptor from donor emission spectra and remove donor spectral bleedthrough. Linear unmixing uses reference spectra generated from λ stacks from single-label specimens.
A, acceptor; FRET, fluorescence resonance energy transfer; qD, quenched donor; QD, quantum dot; uFRET, uncorrected FRET, including spectral bleedthrough and energy transfer levels.