Table 1 ADAPT-3D formulation.
ADAPT-3D Refractive Index Matching Solution (ADAPT:RI) | ||||
|---|---|---|---|---|
Reagent | Amount | Final Concentration | ||
Iohexol | 7.6 g | 25.33% (w/v) | ||
D-Sucrose | 9.0 g | 30% (w/v) | ||
Urea | 7.5 g | 25% (w/v) | ||
n-propyl gallate | 0.150 g | 0.5% (w/v) | ||
1-thioglycerol | 0.3 mL | 1% (v/v) | ||
Iodixanol | To 30 mL | |||
ADAPT-3D Decolorization (ADAPT:DC) | ||||
Reagent | Amount | Final Concentration | ||
CHAPS, (3-((3-cholamidopropyl)dimethylammonio)−1-propanesulfonate) | 100 g | 10% | ||
N-Methyldiethanolamine | 250 mL | 25% | ||
N-Butyldiethanolamine | 50 mL | 5% | ||
1,2-hexanediol | 100 mL | 10% | ||
Water | To 1 L | |||
ADAPT-3D Partial Delipidation Buffer (ADAPT:PDL) | ||||
Reagent | Amount | Final Concentration | ||
Tetrahydrofuran | 5 mL | 50% | ||
1,2-hexanediol | 5 mL | 10% | ||
ADAPT-3D Decalcification Buffer (ADAPT:Decal) | ||||
Reagent | Amount | Final Concentration | ||
EDTA | 150 g | 15% (w/v) | ||
Imidazole | 150 g | 15% (w/v) | ||
N-methyldiethanolamine | 100 mL | 10% (v/v) | ||
N-Butyldiethanolamine | 50 mL | 5% (v/v) | ||
Water | To 1 L | |||
ADAPT-3D Block-and-Stain Buffer (ADAPT:BS) | ||||
Glycine | 3.75 g | 0.1 mM | ||
Tween-20 | 0.833 mL | 0.17% | ||
Triton X-100 | 1.67 mL | 0.33% | ||
Donkey Serum | 1% | 5 mL | ||
Alpaca Serum | 1% | 5 mL | ||
BSA | 1% | 16.67 mL | ||
H2O2 | 1.67 mL | 0.1% | ||
Dimethyl Sulfoxide | 25 mL | 5% | ||
TBS | To 500 mL | |||
ADAPT-3D Fixative (ADAPT:Fix) | ||||
Paraformaldehyde | 4 g | 4% (w/v) | ||
Sodium Hydroxide (*Alternative: Triethanolamine) | pH to 9.0 | |||
1X PBS | To 1 L | |||
Sucrose | To 10% (w/v) | |||
