Pannoramic midi digital section scanner

PI3K (ab86714, Abcam), AKT3 (bs-5146R, Bioss), mTOR (2972s, CST), LC3B (bs-2912R, Bioss), P62 (18420-1-AP, Triple Eagle), western removal buffer of primary antibody and second antibody (P0025, Beyotime), RIPA cell lysate (P0013B, Beyotime), ECL hypersensitive luminescence kit (34095, Thermo), goat anti-mouse IgG secondary antibody (ZB-2305, Zsbio), goat anti-rabbit IgG secondary antibody (ZB-2301, Zsbio), goat anti-rabbit IgG (FITC) (B029, Ebiogo), sheep serum block (B010, Ebiogo), anti-fluorescence quench blocking agent (containing DAPI) (B024, Ebiogo), TRIzol (15596026, Life Technologies), and hematoxylin (BA-4041, BaSO).
EPS 300 electrophoresis instrument (Tanon), VE-180 electrophoresis tank (Tanon), JW-3021HR high-speed refrigerated centrifuge with 6.8cm centrifugal radius (Anhui Jiawen Instrument Equipment), JEM1400 flash transmission electron microscope (Jieou Lu, Beijing), PIKOREAL 96 fluorescence quantitative PCR instrument (Thermo), OD1000+ ultra-micro spectrophotometer (Nanjing Wuyi), CX41 microscope (Olympus), RM2016 Leica microtome (Leica), Pannoramic MIDI digital section scanner (3DHISTECH), and a 1319A digital thermometric indicator(Shanghai TES).

The distal colon tissue was fixed with 4% paraformaldehyde for 24 h, then embedded in paraffin and cut into sections 5 µm thickness for hematoxylin-eosin (HE) staining at Servicebio company (China). The HE staining images were scanned using Pannoramic MIDI Digital Section Scanner (3DHISTECH, Hungary), and the images were analyzed by Case View software. The histological scores of the colon were assessed given a score range from 0 to 4 as previously described parameters: inflammation, depth of inflammation, crypt damage, loss of goblet cells, and thickness of the colon wall (22 (link), 23 (link)).