(A) Whole-body and whole-organ clearing protocols. The whole-organ clearing protocol (left) and the whole-body clearing protocol (right) are shown.

(B) Bright-field images of whole organs (brain, heart, lung, liver, and kidney) from C57BL/6N mouse (13 weeks old, male) after RI adjustment (RI = 1.44, 1.48, and 1.52). Control organs were stocked in PBS buffer (RI = 1.33).

(C) Average parallel transmittance of RI-adjusted brain and lung. Parallel light transmittance around the visible region (450–750 nm) of organs was measured (n = 2). Data represent mean ± SD.

(D) Bright-field images of whole body of C57BL/6N mouse (6 weeks old, male) treated with CUBIC-R. After delipidation and decolorization, adult mouse bodies were treated with PBS buffer (upper) or CUBIC-R (lower).

(E) Whole-body and whole-organ staining protocols. The whole-body staining protocol with PI (left) and the whole-organ staining protocol with RedDot2 or fluorescently labeled primary antibody (right) are shown.

(F) The 3D-reconstituted (3D) body images of PI-stained adult C57BL/6N mouse (10 weeks old, male). Whole-body imaging (WBI), thoracic organ imaging, and abdominal organ imaging are shown.

(G) The 3D-immunostained organ images. The brain from C57BL/6N mouse (15 weeks old, male) was stained with RedDot2 and fluorescein isothiocyanate (FITC)-conjugated anti-α-SMA antibody. 3D (upper) and x-y plane (XY) (lower) images are shown. The white inset was magnified next to the XY image.