Imaging parameters
| Figure | Specimen | Pixel Size (μm) | Z interval (μm) | Slices | Projection | Channels | Excitation (nm) | Emission (nm) | System |
|---|---|---|---|---|---|---|---|---|---|
| 1C | Fixed | 0.41 | 2 | 76 | Maximum | Subtracted | 858 | 470–550, 570–640 | Ti:sapphire laser, 20×/1.0 NA |
| 2A | Fixed | 0.18 | 0 | 1 | N/A | Single, merged | 424–448 (green), 538–562 (magenta), LED (gray) | 460–500 (green), 570–640 (magenta), transmitted light (gray) | Mercury arc lamp and LED, 2.3×/0.57 NA |
| 2B | In vitro | 0.64 | 0 | 1 | N/A | Single, merged | 450–490 (red), LED (gray) | 500–550 (red), transmitted light (gray) | Mercury arc lamp and LED, 10×/0.3 NA |
| 3A | In vitro | 0.58 | 0 | 1 | N/A | Single, merged | 405 (red), 488 (gray) | 430–470 (red), transmitted light (gray) | Diode laser (red), Ar laser (gray), 10×/0.4 NA |
| 3B, left 3 | Clarified | 3.29 | 20 | 51 | Maximum | Merged | 858 | 470–550, 570–640 | Ti:sapphire laser, 20×/1.0 NA |
| 3B, TUNEL | Declarified | 3.29 | 20 | 11 | Maximum | Merged | 1000 | 470–550, 570–640 | Ti:sapphire laser, 20×/1.0 NA |
| 3C | Fixed | 2.49 | 0 | 1 | N/A | Single | 458 | 480–495 | Ar laser, 10×/0.4 NA |
| 3D | Acute slice | 0.82 | 10 | 15 | Sum | Subtracted | 858 | 470–550, 570–640 | Ti:sapphire laser, 20×/1.0 NA |
| 3E | Acute slice | 0.2 | 2 | 21 | Maximum | Merged | 858 | 470–550, 570–640 | Ti:sapphire laser, 20×/1.0 NA |
| 4A, left 6 | Clarified | 3.29 | 20 | 51 | Maximum | Single, merged | 858 | 470–550, 570–640 | Ti:sapphire laser, 20×/1.0 NA |
| 4A, TUNEL | Declarified | 3.29 | 20 | 11 | Maximum | Merged | 1000 | 470–550, 570–640 | Ti:sapphire laser, 20×/1.0 NA |
| 4C | Fixed | 0.82 | 5 | 41 | Maximum | Single, merged | 858 | 470–550, 570–640 | Ti:sapphire laser, 20×/1.0 NA |
| 4D | Fixed | 0.82 | 5 | 41 | Maximum | Single, merged | 858 | 470–550, 570–640 | Ti:sapphire laser, 20×/1.0 NA |
| 4E | Fixed | 0.82 | 10 | 21 | Maximum | Single (row 1), subtracted (row 2) | 858 | 470–550, 570–640 | Ti:sapphire laser, 20×/1.0 NA |
| 4F, left | In vivo | 9.29 | 0 | 1 | N/A | Single | 424–448 | 460–500 | Mercury arc lamp, 1×/0.25 NA |
| 4F, right | In vivo | 9.29 | 0 | 1 | N/A | Single | 424–448 | 460–500 | Mercury arc lamp, 1×/0.25 NA |
| 5A | Clarified | 3.29 | 20 | 51 | Maximum | Merged | 858 | 460–500, 520–550 | Ti:sapphire laser, 20×/1.0 NA |
| 5B | Fixed | 2.27 | 0 | 1 | N/A | Single | 450–490 | 500–550 | Mercury arc lamp, 2.3×/0.57 NA |
| 5C | Fixed | 3.29 | 20 | 14 | Maximum | Merged | 858 | 470–550, 570–640 | Ti:sapphire laser, 20×/1.0 NA |
| 5D | Fixed | 0.26 | 2 | 12 | Maximum | Single, merged | 405 (gray), 561 (magenta) | 415–470 (gray), 570–607 (magenta) | Diode (gray), DPSS (magenta), 40×/1.2 NA |
| 7A | Fixed | 0.82 | 5 | 51 | Maximum | Single, merged | 858 | 470–550 (green), 570–640 (magenta) | Ti:sapphire laser, 20×/1.0 NA |
| 7B | Fixed | 0.82 | 5 | 17 (col 1), 12 (col 2), 3 (col 3) | Maximum | Single, merged | 858 | 470–550 (green), 570–640 (magenta) | Ti:sapphire laser, 20×/1.0 NA |
| 7D | Fixed | 0.55 | 5 | 51 | Maximum | Subtracted | 858 | 470–550, 570–640 | Ti:sapphire laser, 20×/1.0 NA |
Related to Figures 1-5 and 7.