Table 1.
Method-dependent uncertainty ranges of chromatic shifts.
| Origin of Shift | Methods | ||||
|---|---|---|---|---|---|
| Calibration slide (beads) | Calibration slide (biological) | Cross-talk on demand | Replicates on the coverslip | Itself | |
| 3D alignment by Chromagnon | 13–17 (14) | 13–17 (14) | 13–17 (14) | 13–17 (14) | 13–17 (14) |
| Dispersion of samples | 6–440 (100) | 6–30 (15) | 6–700 (40) | 6–30 (15) | 0 |
| Difference in coverslips | 0–1000 (40) | 0–1000 (40) | 0 | 0 | 0 |
| Difference in spherical aberration | 0–225 (100) | 0–225 (100) | 0 | 0 | 0 |
| Mechanical drifts | 0–2000 (100) | 0–2000 (100) | ~0 | ~0 | 0 |
| Total | 20–3682 (354) | 20–3272 (269) | 19–717 (54) | 19–47 (29) | 13–17 (14) |
| Recommended microscopy technique | TIRFM, SMLM | CLSM, STED, SIM | WFM, SMLM | CLSM, STED, SIM | |
Values are typical minimum-maximum (mean) values, expressed in nm, determined from all the data presented in this study. The origins of the shifts depend on the imaging methods and may not be applicable to certain microscopy modalities. Abbreviations: TIRFM, total internal reflection fluorescence microscopy; SMLM, single-molecule localization microscopy; CLSM, confocal laser scanning microscopy; STED, stimulated emission depletion; SIM, structured illumination microscopy; WFM, wide-field microscopy.