Light-sheet microscopy images of aSMA-covered vasculature in human skin and s.c. adipose tissue. (A– E ) Light-sheet microscopy images of cleared and immunostained biopsies of human dermis. Images show (A) PDPN⁺ LYVE-1⁺ LVs, but no PDPN⁺LYVE-1⁻ LVs, and (B) LYVE-1⁺aSMA⁻ LVs and aSMA⁺ BVs. (C–E) Representative images of cleared and immunostained biopsies of human dermis acquired by light-sheet microscopy (n = 6–10 donors per condition). C and D show PDPN⁺aSMA⁻ LVs and aSMA⁺ BVs, and D shows a side view of the skin with the asterisk * indicating the location of the epidermis. Note that D is the same image as the one shown in C (rotated by 90°). 3D views of B–D are provided in Videos 1 and 2. Images in B–D were acquired through the epidermis with a stack height (i.e., penetration depth) of 2.2 and 1.8 mm, respectively. (E) Skin whole mount stained for aSMA and vWF. (I) Overview of the entire skin piece imaged with the superficial and deeper vascular plexus visible, imaged from the side to reveal the two vascular plexi. The asterisk (*) indicates the location of the epidermis. Scale bar: 400 µm. (II) Magnification of the squared angle marked with a dotted line in I, imaged from the bottom of the sample, i.e., from a different orientation. Scale bar: 200 µm. (III) Magnification of the circled area marked with a dotted line in II, zooming in on an aSMA⁺vWF⁻ vessel likely representing a dermal lymphatic collecting vessel. Scale bar: 100 µm. Note that this was the only aSMA⁺vWF⁻ vessel detected in >6 immunostained punches from 6 donors. A 3D view of E is provided in Video 3. (F) Light-sheet microscopy images of cleared and immunostained biopsies of human s.c. adipose tissue. PDPN⁺ or LYVE⁺ LVs (both antibodies detected in the same color) running along larger aSMA⁺ BVs. Representative images of >2/3 (experiments/donors) (A), >3/6 (experiments/donors) (B), 5 (C and D), and >10 (F) experiments are shown. Scale bars: (A) 100 µm, (B) 500 µm, (C and D) 150 µm, (F) 100 µm. Tissue source in light-sheet images shown: breast (A, B, and E), thigh (C and D), and abdomen (F).