表 1. Strengths and weaknesses of major tissue clearing techniques.
主要组织透明化技术优缺点
| Method | Resolution | Permeabilization and delipidation | Preservation of fluorescence | Clearing
time |
| BABB: Benzyl alcohol/benzyl benzoate; iDISCO: Immunolabeling-enabled three-dimensional imaging of solvent-cleared organs; vDISCO: Ultimatethree-dimensional imaging of solvent-cleared organs; Scale: Aqueous reagent that renders biological samples optically transparent; ScaleS: Sorbitol-based optical clearing; CUBIC: Clear, unobstructed brain/body imaging cocktails and computational analysis; SeeNet: 3D visualization and molecular characterization of the entire vascular network; CLARITY: Clear lipid-exchanged acrylamide-hybridized rigid imaging/immunostaining/in situhybridization-compatible tissue hydrogel; PARS: Perfusion-assisted agent release in situ；PACT: Passive clarity technique. | ||||
| Hydrophobic-based tissue clearing | ||||
| BABB | 0.5-2 μm | EtOH (Ethanoal), DCM (Dichloromethane) | Hours | 7 d |
| iDISCO | 0.5-2 μm | Methanol, DCM (Dichloromethane) | Hours | 7-14 d |
| vDISCO | 1-2 μm | Tert-Butanol, THF (Tetrahydrofuran)
DCM (Dichloromethane)/Triton-X100 |
Hours-days | 5 d |
| Hydrophilic tissue clearing | ||||
| Scale | 0.5-2 μm | Urea C, Sorbitol C, DMSO (Dimethyl Sulfoxide) | Days-months | 2-5 d |
| ScaleS | 0.5-2 μm | Urea C, Sorbitol C, DMSO (Dimethyl Sulfoxide) | Days-months | 2-5 d |
| CUBIC | 0.5-2 μm | CUBIC-L (10% Triton-X100 C, 10% Aminoalcohol) | Days | 7-14 d |
| SeeNet | (6.58±1.21) µm | SDC (Sodium deoxycholate) | >6 weeks | 1 week |
| Hydrogel-based tissue clearing | ||||
| CLARITY | 0.5-2 μm | 4% SDS (Sodium dodecylsulfate) | Temperature-dependent:
preserved at 37 ℃ |
1-3 weeks |
| PARS | 1-30 μm | 8% SDS (Sodium dodecylsulfate) | 3 months | 1-4 d |
| PACT | 0.5-12 μm | 8% SDS (Sodium dodecylsulfate) | 3 months | 1-4 d |