Figure 3
CLARITY for whole human organs. (a) CLARITY enables high-resolution and high-quality whole-organ tissue processing and analysis of human fetal tissues. A day 101 fetal sample (top row) can be stained, with Insulin (Ins) and β-tubulin (Tuj1), and imaged through several millimeters of cleared tissue without loss of resolution. Four example z positions spanning the entire pancreas thickness are shown with islets (arrowheads) and nerves/vessels (arrows) observed at each depth. The same tissue can be stained multiple times with different sets of antibodies – Type 4 Collagen (Col4) and Glucagon (Gcg) - (bottom row) with minimal loss of feature resolution. Scale bars = 500 μm whole sample, 100 μm inset. Samples were embedded in A1B1P4 hydrogel and cleared for 2–3 weeks at 60 °C. Antibody removal was performed by clearing for 5–7 days at 60 °C. (b) CLARITY enables automated quantification of structural features, including islet size. In this cohort, there was no significant difference (unpaired t-test) in average islet size, population distributions, or spatial distributions of islet sizes at different developmental ages between d91 and d117. Error: s.e.m. n = 3.
