Dp72 digital camera

All CIN and cancer lesions were examined in a magnified area (×100) for quantification. Images were captured using an Olympus BX50 microscope and a DP72 Olympus digital camera, and cellSens standard 1.5 software (Olympus). We selected the area showing the most severe lesions in each case, and the corresponding epithelial lesions were delineated as ROIs. Both areas of FOXP4‐positive cells and hematoxylin‐stained cells in each ROI were calculated, the ratio of the FOXP4‐positive and hematoxylin‐stained areas was defined as the FOXP4‐positive rate, and values were normalized using National Institutes of Health (NIH) ImageJ software, as described previously.⁹

The W12 cells transfected with FOXP4‐shRNA and NC‐shRNA were cultured on glass chamber slides (Lab‐Tek® II, Themo Fisher Scientific). A slide with 80% confluent cells was fixed in 4% paraformaldehyde and stained by Hoechst® 33342 (Themo Fisher Scientific). The stained nuclei were scanned using a confocal laser scanning microscope (LSM510; Carl Zeiss) and sagittal images were reconstructed. Horizontal images (×100) were captured using an Olympus BX50 microscope and a DP72 Olympus digital camera, and cellSens standard 1.5 software (Olympus). The sites showing overlapping nuclei were defined as cell‐overlapping zones, and the total area of overlapping zones was calculated using NIH ImageJ software.

The CRC tissues and matching normal colon tissue specimens were embedded in paraffin wax and sectioned at about 5-μm thickness with a microtome (Leica Microsystems GmbH, Wetzlar, Germany). The tumorareas in IHC slides were defined by staining each section with hematoxylin and eosin stains. The array tissue blocks were obtained as described elsewhere 45 (link) and each slide was incubated in a hot air oven at 60°C for 15-20 minutes, followed by incubation with a primary TLR-6 antibody (Santa Cruz Biotechnology, CA, USA) at a 1/100 dilution. The other steps and analyses for IHC were conducted as described by Semlali et al. 9 (link). The immuno-stained slides were analyzed with an Olympus BX51 light microscope (Olympus America Inc., Center Valley, PA, USA) equipped with aDP72 Olympus digital camera with powers 400×.
IHC analysis was performed by using points (0 to 4) according to the level and range of the color in each condition: 0 point corresponds to no positive color; 1 point corresponds to <20% positive staining; 2 points means a positive staining between 21 50%; 3 points corresponds to , 51-75% positive staining; and 4 points means >75% positive staining.

After the 4-wk restriction of food intake, the rats were killed under anesthesia, and the bilateral ovaries were removed and fixed by 10% formalin. Ovaries were embedded in paraffin in parallel to their long axis. Four-micrometer-thick tissue slides containing the broadest ovarian tissue section were prepared from paraffin-embedded ovary block and stained by hematoxylin-eosin staining. Images were captured using an Olympus BX50 microscope, a DP72 Olympus digital camera, and CellSens standard 1.5 software (Olympus). Initial selections of follicles (mean diameter, >250 µm) and corpora lutea (>600 µm) were performed based on their sizes using low-magnification digital photographs and the corresponding scale bars (Figure 2A). Next, the healthy growing follicles and corpora lutea were independently evaluated under a microscope by 3 gynecologic endocrinologists (14 (link)) without any information about the origin of each slide (Figure 2B and C), and then their numbers were finally determined (Figure 2A, asterisks). If the judgment of health conditions differed, the less favorable evaluation was selected, as reported previously (15 (link), 16 ).