Digital sight ds fi2 camera

The specimens were processed for H&E staining and IHC as described [8 (link), 9 (link)], with antibodies for NHERF1 1:3200 (Thermo/Fisher, Waltham, MA, USA), moesin 1:100 (3150, Cell Signaling Technology, Danvers, MA), NF2 C-terminal 1:800 (C18) (see also [35 (link)] (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and EMA 1:400 (Dako, Carpinteria, CA) on a Leica automated IHC platform (Leica Biosystems, San Diego, CA, USA). The protocol for NHERF1 immunostaining was subsequently validated for clinical use according to the College of American Pathologists (CAP) laboratory quality assurance guidelines on a Ventana Benchmark Ultra platform (Roche/Ventana Medical Systems Inc., Tucson, AZ, USA) at 1:2000 dilution. Progesterone receptor (1E2) rabbit monoclonal antibodies (Roche/Ventana) IHC was used on selected cases. Cytology preparations were obtained from fresh specimens and processed by H&E manual staining during intraoperative diagnosis. Images were acquired at various magnifications with an Aperio Scanscope CS2 whole slide image system (Leica Biosystems) or with a Nikon Eclipse Ci microscope equipped with a Nikon Digital Sight DS-Fi2 camera (Nikon Instruments Inc., Melville, NY, USA), by using the Nikon NIS Elements 4.51.00 program.

Human lung tissues collected from LoM were fixed in 10% formalin, paraffin embedded, and cut into 5 µm sections which were mounted onto Superfrost Plus slides (Fisher Scientific). Tissue sections were incubated at 60°C for 1 h, deparaffinized with xylene (2 × 3 min) and graded ethanol (100% 2 × 3 min, 95% 1 × 3 min, 80% 1x 3min, 70% 1 × 3 min), and stained with hematoxylin followed by eosin. Tissue sections were then mounted and imaged on a Nikon Eclipse Ci microscope using Nikon Elements BR software (version 4.30.01) with a Nikon Digital Sight DS-Fi2 camera. Brightness, contrast and white balance were adjusted on whole images in Adobe Photoshop (CS6).

Formalin-fixed paraffin-embedded (FFPE) sections from brain tumor resection and lung needle biopsy specimens were stained with hematoxylin-eosin (H&E). Images were acquired with a Nikon Eclipse Ci microscope equipped with a Nikon Digital Sight DS-Fi2 camera (Nikon Instruments Inc., Melville, NY), as previously described [10 (link)]. IHC was performed with clinically validated antibodies on a Ventana Benchmark Ultra platform (Roche/Ventana Medical Systems Inc., Tucson, AZ) [10 (link)]. The primary antibodies were: glial fibrillary acidic protein (GFAP) (EP672Y), Olig-2 (387 M-15) (Ventana/Cell Marque, Rocklin, CA), p53 (DO-7), Ki-67 antibody (30–9) (Roche/Ventana Medical Systems Inc.), IDH1-R132H (DIA-H09, Dianova, Hamburg, Germany) and NHERF1/EBP50 (Thermo/Fisher, Waltham, MA). The automated Ki-67 proliferation index was performed by using the Nikon NIS Elements 4.51.00 program set up with an object-count algorithm for recognition of differentially labeled nuclei, as previously described [32 (link)].

The breads were examined microscopically in order to examine potential structural differences between the breads. Bread samples were embedded as described by Johansson et al. [24 (link)] and the embedded samples were cut into 2 µm thick sections with an ultra-microtome (Leica EM UC6, Leica, Vienna, Austria). Sections were stained with Fast Green/Lugol’s solution or Calcofluor White. Fast Green colored proteins green while Lugol’s solution colored starch purple/violet making it possible to distinguish between amylopectin rich areas (beige/brown), and amylose (blue). When stained by calcofluor, cell walls rich in β-glucan appeared blue when examined in exciting light (excitation, 400–410 nm; emission, 455 nm). The stained sections were examined using a Nikon Eclipse Ni-U microscope (Nikon Instruments Inc., New York, NY, USA) and the images were captured with a Nikon Digital Sight DS-Fi2 camera (Nikon Instruments Inc., New York, NY, USA) and processed with the software NIS-Elements BR (Nikon Instruments Inc., New York, NY, USA).