Figure 7.

Increasing delipidation time from 20 h to 68 h dramatically improves tissue clearing and imaging in aged adult mouse tissues

Lightsheet images of kidney (A and B), heart (C and D), ovary (E), eye (F) and spleen (G) from 5 month old mice following perfusion with far red fluorescent lectin to label blood vessels. Kidney and heart samples were delipidated for 20 h (A and C) and then imaged following RI matching. These exact same tissues were then subjected to an additional 48 h of delipidation prior to a second round of RI matching and imaging (B and D). The kidney sample delipidated for 20 h (A) does not show any discernable vasculature, indicating that this incubation time is not adequate for tissue clearing of 5 month old kidney. After an additional 48 h of delipidation, the vasculature of the 5 month old kidney (B) is evident. The heart after 20 h of delipidation (C) shows clear signal in the vasculature, although some areas lack crisp signal. However, an additional 48 h of delipidation adds detail previously undetected with the 20 h delipidation and improves the signal to noise ratio. 20 h of delipidation is adequate for the ovary (E) and eye (F), whereas the spleen requires an additional 48 h of delipidation to observe optimal signal. Scale bars in A1–D1 = 500 μm; A2–E1 = 200 μm; F1 = 300 μm; G1 = 700 μm; E2 = 70 μm; F2 = 100 μm; G2 = 200 μm.