Pannoramic midi and pannoramic viewer

Human Pancreatic tissue was fixed with 10% formalin, followed by dehydration, paraffin embedding, and sectioning at 3 μm. Sections were then stained according to the protocol in one of our earlier studies [5 (link)]. After antigen retrieval of In brief, after deparaffinization, the paraffin sections were blocked with blocking buffer (10% FBS, 0.1% Triton X-100) for 30 min at 37°Cfollowed by sequential incubations with anti-NKX6.1 (1:500, NBP1–49672, Novus, USA) and anti-insulin (1:200, ab7842, Abcam, USA) at 4 °C overnight and secondary antibodies (Alexa Fluor 488 AffiniPure Goat Anti-Guinea Pig IgG H&L, 1:200, 106–545-003, Jackson Immunoresearch Laboratories and Molecular Probes, USA; Rhodamine (TRITC) AffiniPure Goat Anti-Rabbit IgG H&L, 1:100, 111–025-003, Jackson Immunoresearch Laboratories and Molecular Probes, USA) for 30 min at 37 °C, and then counterstained with DAPI. Pancreatic tissues were scanned by Pannoramic MIDI and Pannoramic Viewer (3DHistech).
Quantifications were performed in a blinded fashion using the CytoNuclear count function of the Image Pro Plus software (Media Cybernetics, Silver Spring, Maryland). Cells with a strong nucleic NKX6.1 immunreactivity in islet were described as nucleus NKX6.1⁺ cells, and cells with cytoplasmic or no NKX6.1 immunreactivity were described as nucleus NKX6.1⁻ cells.