Optiview dab ihc detection

Immunohistochemistry (IHC) was performed using patient colon tissues, which were fixed with 4% paraformaldehyde and embedded in paraffin. Immunostaining was performed using the BenchMark XT automatic immunostaining device (Ventana Medical Systems, Inc., Oro Valley, AR, USA) and OptiView DAB IHC Detection (Ventana Medical Systems, Inc.). Tissue sections (4 μm) were transferred to salinized, charged slides and incubated at room temperature and 65°C. After antigen retrieval for 64 min, the sections were incubated on a fully automated immunostainer with anti-iNOS antibody (Abcam, Cambridge, UK;) for 32 min. Tissue section slides were then counter-stained with 4′,6-diamidino-2-phenylindole (DAPI). All image-staining patterns of the slides were acquired using an OptiView DAB IHC Detection Kit (Ventana Medical Systems, Inc.). Furthermore, all images were acquired using a x20 objective lens.

Standardized automated immunohistochemistry is performed using the Benchmark Ultra IHC platform with OptiView DAB IHC detection (Ventana Medical Systems) per manufacturer specification. Primary antibody (anti-phospho-TDP-43 (pS409); Cat#: TIP-PTD-P03, Cosmo Bio) [22 (link)] is incubated at 1/1000 for 32 minutes at 36°C following antigen 40 minutes retrieval without amplification.