Lillie mayer s hematoxylin

Segments of human jejunum, tHIO and tHIO+S grafts were fixed overnight in 4% paraformaldehyde (PFA), processed and embedded in paraffin. Sections of 5µm thickness were deparaffinized and either stained immediately with a histological kit (Pentachrome, Newcomer Supply) or subjected to antigen retrieval, and stained. For both primary and secondary antibodies, incubations took place at 4°C overnight in 1% bovine serum albumin in phosphate buffered saline (PBS). Antibodies, their respective dilutions are listed in Table S1. The Vectastain ABC system (Vector Laboratories) was used for amplification. The following kits were used for signal detection: diaminobenzidine substrate kit, Immpact SG substrate kit, and Vector Red substrate kit (Vector Laboratories). Lillie-Mayer’s Hematoxylin (Dako North America, Inc.) or Nuclear Fast Red (PolySciences, Inc.) was used as a counterstain. Images were acquired using a Nikon Eclipse Ti microscope and analyzed using Nikon Elements Imaging Software (Nikon).