FIGURE 2.

Summary of optical clearing methods applied to the ovary. (A) Outline of organic solvent-based and aqueous-based clearing approaches. Despite the use of different reagents in two different (BABB and iDISCO) organic solvent-based clearing methods, both protocols involve dehydration, delipidation, and a final RI matching step. Aqueous-based approaches (CUBIC and ScaleA2) use high concentrations of detergents to remove lipids, while hydrogel embedding of the sample in CLARITY protocol maintains tissue and protein structure before incubation with high concentration of detergents to remove lipids. (B) Comparison of different methods based on experimental specifications. Studies used combined approaches are placed spanning the columns for iDISCO and CUBIC or ScaleA2 and CLARITY in the reference column. BABB, benzyl alcohol and benzyl benzoate; iDISCO, immunolabeling-enabled three-dimensional imaging of solvent-cleared organs; CUBIC, clear, unobstructed brain/body imaging cocktails and computational analysis; CLARITY, clear lipid-exchanged acrylamide-hybridized rigid imaging/immunostaining/in situ-hybridization-compatible tissue hydrogel; RI, refractive index.