Table 1 Staining protocol used in Experiments 1 and 2
Process | Step | Solution | Duration (m:s) | Â | Process | Step | Solution | Duration (m:s) |
|---|---|---|---|---|---|---|---|---|
Dewaxing | 1 | Xylene (wash 1) | 3:00 | Â | Staining | 14 | Running tap water | 1:00 |
2 | Xylene (wash 2) | 3:00 |  | 15 | Scott’s Tap Water | 2:00 | ||
3 | Xylene (wash 3) | 3:00 | Â | 16 | Running tap water | 1:00 | ||
4 | Xylene (wash 4) | 3:00 |  | 17 | Aqueous Eosin Y (1%) | 0:15 − 6:00 (see SI Table 2) | ||
Rehydration | 5 | 100% ethanol (wash 1) | 3:00 | Â | 18 | Running tap water | 1:00 | |
6 | 100% ethanol (wash 2) | 3:00 | Â | Dehydration | 19 | 100% ethanol (wash 1) | 0:30 | |
7 | 100% ethanol (wash 3) | 3:00 | Â | 20 | 100% ethanol (wash 2) | 1:00 | ||
8 | 100% ethanol (wash 4) | 3:00 | Â | 21 | 100% ethanol (wash 3) | 5:00 | ||
9 | 75% ethanol | 3:00 | Â | 22 | 100% ethanol (wash 4) | 5:00 | ||
10 | 50% ethanol | 3:00 | Â | Clearing | 23 | Xylene (wash 1) | 3:00 | |
11 | 25% ethanol | 1:00 | Â | 24 | Xylene (wash 2) | 3:00 | ||
12 | Running tap water | 2:00 | Â | 25 | Xylene (wash 3) | 3:00 | ||
Staining | 13 | Mayer’s Haematoxylin | 0:15 − 6:00 (see SI Table 2) |  | Mounting | 26 | DPX mountant | - |