Mc120 hd camera

Formalin-fixed and paraffin-embedded tissues were processed and sectioned according to routine protocols. Heat mediated antigen retrieval was used prior to all staining procedures. Tissues were incubated with vimentin antibody (1:200 dilution, clone D21H3, Cell Signaling Technology) or S100A9 antibody (1:100 dilution, provided by Dr. Philippe Tessier at Laval University) overnight at 4 °C. Antigen was detected using the EnVision+ Dual Link System (Dako) and counterstained with hematoxylin. Images were taken using a Leica DM4000B microscope and a Leica MC120 HD camera with a 40× objective.

Plant samples were submerged in ice-cold 90% (v/v) acetone for 5 min. After washing with distilled water, samples were vacuum infiltrated with staining solution (100 mM sodium phosphate buffer pH 7.0, 10 mM EDTA, 0.1% [v/v] Triton X-100, 0.5 mM K₃Fe(CN)₆, 0.5 mM K₄Fe(CN)₆, 2 mM 5-bromo-4-chloro-3-indolyl-ß-glucuronide, and 20% [v/v] methanol), and then the infiltrated samples were incubated at 37 °C for 1 h. Stained samples were submerged in 70% (v/v) ethanol and observed using a stereomicroscope (M165FC, Leica) with a MC120 HD camera (Leica).

At the experimental endpoint, mice were anesthetized with isoflurane, and blood collected using cardiac puncture. A midline incision was made in the abdominal cavity to isolate and excise the colon, and colon length was measured. The distal colon was collected and fixed in 10% formalin for 24 h, paraffin-embedded, cut into 4 µm sections and stained using hematoxylin and eosin (H&E). Images of the colon sections were taken at 4x magnification using the Leica DM LB widefield microscope and MC120 HD camera (Leica), and scored in a blinded manner. Parameters for histology scoring were as previously described (24 (link)). Following collection of the distal colon for histology, the rest of the colon was used for flow cytometric analysis or sectioned into two parts for myeloperoxidase (MPO) assay (middle section) and RNA isolation (proximal section).

At 24 hr after MCAO, a section of the distal colon was collected for haematoxylin and eosin (H&E) staining. For each sample, two representative images of colonic crypts at 100× magnification were captured using the Leica DM LB widefield microscope and MC120 HD camera (Leica) for histological scoring in a blinded manner. Parameters for histology scoring were previously established (Shen et al., 2018) and detailed in Data S1. Total overall score indicates the degree of colonic pathology after MCAO or sham operation.

Microscopic inspection for signs of uniform and local corrosion was performed with a VHX-S650E digital microscope (Keyence Corporation, Osaka, Japan) with 20–100× magnification at DTU and a Leica MC 120 HD camera at FT. Microstructure examination was performed on cut sections of selected nails, followed by mounting in resin and subsequent grinding and polishing down to 1 µm diamond polishing. After etching in Vogels Sparbeize etchant, the microstructure was examined using a Leica DMI 5000 M metallographic microscope.

At the experimental endpoint, mice were anesthetized with isoflurane, and blood collected using cardiac puncture. A midline incision was made to isolate and excise the colon, and colon length was measured. The distal colon was collected and fixed in 10% formaldehyde for 24 h, paraffin-embedded, cut into 4 µm sections, and stained using hematoxylin and eosin (H&E). Images of the colon sections were taken at 4× magnification using the Leica DM LB widefield microscope and MC120 HD camera (Leica), and scored following blinding and randomization. Parameters for histology scoring includes the level of tissue involvement (0: none, 1: mucosa, 2: mucosa and sub-mucosa, 3: sub-mucosa-transmural, 4: transmural), level of inflammation (0: none, 1: mild, 2: moderate, 3: severe, 4: severe with gastrointestinal-associated lymphoid tissue involvement), involvement of crypt and epithelium (0: none, 1: surface, 2:2/3 basal, 3: crypt and goblet cell loss, 4: crypt and goblet cell destruction with hyperplasia, surface epithelial destruction), and level of lamina propria/sub-mucosa edema (0: none, 1: mild, 2: moderate, 3: moderate-severe, 4: severe). Following collection of the distal colon for histology, the rest of the colon was used for flow cytometric analysis, or sectioned into two parts for myeloperoxidase assay (middle section) and RNA isolation (proximal section).

At 24 hr after MCAO, a section of the distal colon was collected for Periodic acid‐Schiff reagent (PAS) staining according to standard protocols. For each sample, two representative images of colonic crypts at 100× magnification were captured using the Leica DM LB widefield microscope and MC120 HD camera (Lecia). The number of goblet cells per colonic crypt was quantified and presented as number of cells per mm².