Figure 2.

Modified CUBIC protocol allows for imaging of pigmented and albino eyes

(A and B) Albino ROSA^mT/mG mouse eyes cleared using (A) iDISCO and (B) CUBIC clearing techniques. Organic solvent-based methods, such as iDISCO, result in significant shrinkage of the retina consequently causing the retina to collapse inwards and adversely affect the 3D tissue morphology. Dotted white lines in the 2D cross-section of the entire eye demonstrate the morphological change of the retina. The retina is shaped more like a ‘V’ after iDISCO compared to a more circular shape after CUBIC clearing. Magnified regions are from the white boxed area in the 2D cross-section. Scale bars, 2D cross section: 500 μm, magnification: 100 μm.

(C and D) (C) Pigmented and (D) albino mouse eyes before (C and D) and after (C′ and D′) applying our clearing protocol. 1 mm grid. ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer; on, optic nerve. Dashed lines in magnification indicate layers of the retina.