60°C for 10 min. We treated the samples with protease III (ACD, Cat# 322337) for 30 min at 40°C and incubated them with probes for 2 hr. After hybridization with AMP 1, 2 and 3, we incubated the slides with different HRP channels and fluorophores, including Opal 520 (PerkinElmer, Cat# FP1487A), Opal 570 (PerkinElmer, Cat# FP1488A) and Opal 650 (PerkinElmer, Cat# FP1496A). Lastly, we used Trueblack (Biotium, Cat# 23007) to quench Lipofuscin autofluorescence for 45 s at room temperature and counterstained every slide with DAPI before mounting with Prolong Gold Antifade Mountant (Life technologies, Cat# P36930).
