Standardized H&E Staining of Mouse Tissues

H&E staining of mouse organs and tumours was performed by the staff at the Anatomical Pathology Laboratory Services located at The Royal Children’s Hospital, Melbourne, Australia according to the standard H&E protocol as described previously [28 (link)]. Paraffin embedded tissues were sectioned at 4 µm thickness, deparaffinised, rehydrated and stained for 3 min with Haematoxylin (Australian Biostain Pty Ltd, Traralgon, VIC, Australia). Slides were rinsed with 0.25% Acid Alcohol and Scott’s Tap water Substitute. Sections were stained with Eosin (Amber Scientific, Midvale, WA, Australia) for 2 min before final rinsing with absolute alcohol and xylene. The metastatic development was assessed by a qualified pathologist at The Royal Women’s Hospital (Melbourne, VIC, Australia). Histological images of stained organs and tumours were taken using Evos FL Auto 2 microscope (Thermo Fisher Scientific).