| Sample . | Error . | References for correction protocols . |
|---|---|---|
| (a) Fluorescent microspheres (beads) that are below the diffraction resolution limit of the imaging system | Optical aberrations | Hiraoka et al., 1990; Goodwin, 2013 |
| (b and c) Multi-wavelength beads below the diffraction resolution limit of the imaging system | Channel registration | Hibbs et al., 2006; Spiering et al., 2013; Wolf et al., 2013 |
| (d) Stage micrometer | In magnification/pixel size | Wolf et al., 2013 |
| Flatfield slide | Nonuniform illumination | Model, 2006 |
| Single-labeled biological sample | Bleed-through | Spiering et al., 2013 |
| Bolte and Cordelières, 2006 | ||
| Stable biological sample | Photobleaching | Bancaud et al., 2010 |
| Unlabeled biological sample | Autofluorescence | Hibbs et al., 2006 |
| Sample . | Error . | References for correction protocols . |
|---|---|---|
| (a) Fluorescent microspheres (beads) that are below the diffraction resolution limit of the imaging system | Optical aberrations | Hiraoka et al., 1990; Goodwin, 2013 |
| (b and c) Multi-wavelength beads below the diffraction resolution limit of the imaging system | Channel registration | Hibbs et al., 2006; Spiering et al., 2013; Wolf et al., 2013 |
| (d) Stage micrometer | In magnification/pixel size | Wolf et al., 2013 |
| Flatfield slide | Nonuniform illumination | Model, 2006 |
| Single-labeled biological sample | Bleed-through | Spiering et al., 2013 |
| Bolte and Cordelières, 2006 | ||
| Stable biological sample | Photobleaching | Bancaud et al., 2010 |
| Unlabeled biological sample | Autofluorescence | Hibbs et al., 2006 |
We used (a) Molecular Probes FluoSpheres; (b) for high-resolution imaging, Invitrogen TetraSpeck Microspheres, 0.1 µm; (c) for low-resolution imaging, Invitrogen FocalCheck Beads, 6 µm or 15 µm; and (d) MicroScope World, 25 mm KR812.