Microphoto fxa microscope

Frozen sections (8 μm) of aortic sinus lesions were prepared, and stained for neutral lipid accumulation in lesion sections by Oil red O staining. Collagen content was measured by staining with Masson's trichrome staining kit (HT15‐1KT; Sigma–Aldrich). Necrotic core area was quantified by assessing unstained area in plaques stained with Masson's trichrome. Nuclei for all immunohistochemistry stains were counterstained using Gill's Hematoxylin. Immunohistochemistry stains were visualized with a Nikon Microphoto‐Fxa microscope (Nikon) equipped with an FX‐35‐DX digital camera (Nikon). Images were captured using 4x/0.13 objective lens using the ACT‐2U imaging software (Nikon). Image quantification was performed using Image Pro Plus software. For en face analysis, descending aortas were fixed in 10% formalin, trimmed of fat, stained with Oil red O, opened longitudinally, pinned out on black wax, and digitally photographed, as described.11 Images were captured using a dissecting photomicroscope. Image quantification for en face analysis was performed using Biopix software (Gothenburg, Sweden).

All patients data presented in this work were collected under MDACC institutional review board (IRB)-approved lab protocolLab06-0526. Al the patients provided written informed consent prior to surgical resection. Immunohistochemical staining was performed on formalin-fixed and paraffin-embedded (FFPE) tumor patient sections after deparafinization, antigen retrieval and blockade of endogenous peroxidase activity and total proteins. The primary antibodies diluted in the blocking buffers were added overnight at 4°C for DR4 (BD-Biosciences) and DR5 (BD-Biosciences). Sequentially, the slides were washed and incubated with the secondary antibody. Slides were then developed with 3,3’-diaminobenzidine tetrahydrochloride substrate that includes horseradish peroxidase enzyme and hematoxylin was used for counter staining. Staining was evaluated and scored by HMA. Photomicrographs were captured using a Nikon Microphoto FXA microscope (Nikon Instruments), an Olympus DP70 camera (Olympus America; Jupiter, FL), and the QCapture Suite Plus software (QImaging; Surrey, British Columbia, CA).