Table 1.
Comparison of UbasM properties with other optical clearing methods. The table summarizes general properties and performance indicators for UbasM and other major published clearing methods. These methods are categorized mainly into aqueous solution-/organic solvent-based and hydrogel embedding methods. To evaluate clearing efficiency, mm-thick adult mouse brain slice and 4-week old mouse hemisphere were used as small (~mm) and large samples (~cm). SeeDB was only demonstrated to embryos and young mice and the resulting sample transparency is insufficient for one-photon imaging of the adult organ. The complex uDISCO processing (i.e. 8–9 steps) required at least 15 hours30. Conclusions were evaluated from experiments performed in this paper (indicated by bold text) and also based on published information. (RT – room temperature; N.T. – not tested)
| Method (Ref) | Agents | Clearing Performance | Handling | Incubation Temp. | |||||
|---|---|---|---|---|---|---|---|---|---|
| Main (Detergent) | Clearing Mode (steps) | Capability | Efficiency | FP preservation | DiI compatibility | ||||
| Slice (~mm) | Hemisphere (~cm) | ||||||||
| Organic solvent-based methods | |||||||||
| BABB (ref.24) | Benzyl alcohol Benzyl Benzoate | Replacing water with organic solvents (5–6 steps) | Very Strong | Slow >4.5 hr | Very Fast 3–7 d | Very poor | Poor | Needs Care | RT |
| 3DISCO (ref.25) | Benzyl Ether | Replacing water with organic solvents (6 steps) | Very Strong | Slow >3.5 hr | Very Fast 3–7 d | Poor | Poor | Needs Care | RT |
| iDISCO (ref.40) | Benzyl Ether | Replacing water with organic solvents (6 steps) | Very Strong | Slow >3.5 hrr | Very Fast 3–7 d | Poor | Poor | Needs Care | RT |
| uDISCO (ref.30) | BABB(4): DBE(1) | Replacing water with organic solvents (8–9 steps) | Very Strong | Very Slow>15 hr | Very Fast 3–7 d | Medium | Poor | Needs Care | > 26 °C |
| Aqueous solution-based methods | |||||||||
| Scale (ref.31) | Urea Glycerol (0.1%Triton X-100) | Water-based chemical agents (1–2 steps) | Medium | Very Slow >15 hr | Very Slow months | Excellent | N.T. | Very Easy | RT |
| SeeDB (ref.34) | Fructose | Sugar solutions (6 steps) | Weak | Slow >2 hr | — | Excellent | Good | Easy (viscous) | 37 °C |
| CUBIC (ref.32) | Urea Amino-alcohol (Triton X-100 15%, Reagent-1 0.1%, Reagent-2) | Detergent-based chemical agents (3 steps) | Strong | Fast 1–2 hr | Fast 7–12 d | Excellent | Poor | Very Easy | RT (or 37 °C) |
| ScaleS (ref.33) | Urea Sorbitol (0.2% Triton X-100) | Water-based chemical agents (5 steps) | Strong | Medium >2 hr | Medium 2–4 weeks | Excellent | Good (Poor for SQ(5)) | Very Easy | 37 °C |
| UbasM | Urea Amino-sugar (0.2% Triton X-100) | Water-based chemical agents (3 steps) | Strong | Fast 1–2 hr | Fast 7–12 d | Excellent | Good | V ery Easy | RT (or 30 °C) |
| Hydrogel embedding methods | |||||||||
| CLARITY (ref.37) | SDS Boric acid (SDS 4%) | Hydrogel embedding +removal of lipids with electrophoresis | Very Strong | - N.T. | Medium 2–4 weeks | N.T. | N.T. | Complicated Needs device Careful | 37 °C |
| PACT (ref.38) | SDS Histodenz (SDS 8%) | Hydrogel embedding + removal of lipids with 8% SDS | Very Strong | - N.T. | Medium 2–4 weeks | Medium | Poor | Careful | 37 °C |