Cubic
Comprehensive guide
Protocol for Imaging and Analysis of Mouse Tumor Models with CUBIC Tissue Clearin. Article.
Compleate protocol for Mouse Tumor Models.
Whole-mouse Clearing
Mouse whole-Body Clearing (without decalcification process)
Application: Whole Adult Mouse
- Pre-Treatment: 200 mL 50% CUBIC-L at 37°C O/N
- Delipidation: 200 mL CUBIC-L at 37°C for 5 days
- (CUBIC-L refreshed on days 1, 2, and 4)
- Delipidation has reached its endpoint when:
- Partial transparency is acheived
- CUBIC-L remains colorless after incubation with sample
- Delipidation has reached its endpoint when:
- (CUBIC-L refreshed on days 1, 2, and 4)
- Pre-Treatment: 200 mL 50% CUBIC-R+(M) at RT O/N
- RI Matching: 200 mL CUBIC-R+(M) at RT O/N
- Reagent Totals (for a 12 cm x 8 cm x 6 cm container):
- CUBIC-L : 700 mL
- CUBIC-R+(M) : 300 mL Delipidation has reached its endpoint when:
- Reagent Totals (for a 12 cm x 8 cm x 6 cm container):
†Use a container that allows you to submerge your whole specimen.
†For nuclear staining, we recommend using a solution of 30 μg/mL propidium iodide (PI) and 1.5M NaCl in PBS.
FA: formaldehyde, O/N: overnight, PFA: paraformaldehyde, RT: room temperature.
Whole-organ
Decalcification overview
Decalcification (Whole Mouse).
Decalcification + Cubic-HV
Process outline:
Process details:
(see cubic_hv section)