Table 1.
Comparison of decalcification efficiency and clearing time of the rodent bone tissues in this study.
| Bone clearing protocol | Bone CLARITY | Bone-mPACT + | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Decalcification | 10% EDTA | 20% EDTA | Calci-Clear Rapid | 5% Nitric acid | 10% Formic acid | |||||||
| Clearing condition | Decalcification time | Clearing time | Decalcification time | Clearing time | Decalcification time | Clearing time | Decalcification time | Clearing time | Decalcification time | Clearing time | ||
| Tm: 4 °C | 8S | Tm: 45 °C | 8S10C+ | Tm: 45 °C | 8S10C+ | Tm: RT | 8S10C+ | Tm: RT | 8S10C+ | |||
| Mouse | Head (+Brain) | ±96 h (n = 2) | ±1020 h (n = 2) | ±96 h (n = 3) | ±960 h (n = 3) | ±10 h (n = 3) | ±960 h (n = 3) | 8 h (n = 3) | ±720 h (n = 3) | - | - | |
| Lumbar Vertebrae | - | - | ±96 h (n = 5) | ±72 h (n = 5) | ±8 h (n = 5) | ±72 h (n = 5) | - | - | - | - | ||
| Fore limb | Scapula | - | - | ±96 h (n = 5) | ±48 h (n = 5) | ±6 h (n = 5) | ±48 h (n = 5) | - | - | - | - | |
| Humerus | - | - | ±96 h (n = 5) | ±72 h (n = 5) | ±6 h (n = 5) | ±72 h (n = 5) | ±1 h (n = 5) | ±72 h (n = 5) | ±8 h (n = 5) | ±72 h (n = 5) | ||
| Radius-Ulna | - | - | ±96 h (n = 5) | ±72 h (n = 5) | ±6 h (n = 5) | ±72 h (n = 5) | ±1 h (n = 5) | ±72 h (n = 5) | ±8 h (n = 5) | ±72 h (n = 5) | ||
| Hind limb | Femur | - | - | ±96 h (n = 5) | ±96 h (n = 5) | ±6 h (n = 5) | ±96 h (n = 5) | - | - | - | - | |
| Tibia-Fibula | ±96 h (n = 5) | ±120 h (n = 5) | ±96 h (n = 5) | ±96 h (n = 5) | ±6 h (n = 5) | ±96 h (n = 5) | ±1 h (n = 5) | - | ±8 h (n = 5) | - | ||
| Foot | - | - | ±96 h (n = 5) | ±96 h (n = 5) | ±6 h (n = 5) | ±96 h (n = 5) | - | - | - | - | ||
| Rat | Lumbar Vertebrae | - | - | ±168 h (n = 5) | ±240 h (n = 5) | ±2 h (n = 5) | ±240 h (n = 5) | - | - | - | - | |
| Fore limb | Scapula | - | - | ±168 h (n = 5) | ±168 h (n = 5) | ±8 h (n = 5) | ±168 h (n = 5) | ±5 h (n = 5) | ±240 h (n = 5) | ±40 h (n = 5) | ±240 h (n = 5) | |
| Humerus | - | - | ±168 h (n = 5) | ±360 h (n = 5) | ±10 h (n = 5) | ±360 h (n = 5) | ±6 h (n = 5) | ±360 h (n = 5) | ±42 h (n = 5) | ±288 h (n = 5) | ||
| Radius-Ulna | - | - | ±168 h (n = 5) | ±240 h (n = 5) | ±10 h (n = 5) | ±240 h (n = 5) | ±6 h (n = 5) | ±360 h (n = 5) | ±42 h (n = 5) | ±240 h (n = 5) | ||
| Hind limb | Hip Bone | - | - | ±168 h (n = 5) | ±240 h (n = 5) | ±10 h (n = 5) | ±240 h (n = 5) | ±8 h (n = 5) | ±480 h (n = 5) | ±45 h (n = 5) | ±240 h (n = 5) | |
| Femur | - | - | ±168 h (n = 5) | ±240 h (n = 5) | ±12 h (n = 5) | ±480 h (n = 5) | ±8 h (n = 5) | ±600 h (n = 5) | ±45 h (n = 5) | ±360 h (n = 5) | ||
| Tibia-Fibula | - | - | ±68 h (n = 5) | ± 240 h (n = 5) | ±12 h (n = 5) | ±480 h (n = 5) | ±8 h (n = 5) | ±600 h (n = 5) | ±45 h (n = 5) | ±360 h (n = 5) | ||
8S: 8% SDS in 0.1 M PBS; 8S10C + : 8% SDS, 10% SDC and 0.5% α-thioglycerol in 0.1 M PBS.