Dp70 ccd camera

Two chromosome painting probes for the STK39 gene (on the long arm) and GNAQ gene (on the short arm) located on metacentric Chromosome 2 of the Tibetan sheep assembly were produced by PCR and labeled with TexasRed-dUTP and FITC-dUTP through nick translation. The information for the PCR primers and the length of amplified DNA fragments was detailed in Supplemental Table S26. In FISH, metaphase spreads were first permeabilized using Triton X-100 for 10 min and rinsed in PBS for 5 min, treated with pepsin at room temperature for 5 min, rinsed in PBS for 5 min twice and in 70% ethanol for 30 sec. After the slides were air-dried and dehydrated with a gradient of 70%, 85%, and 100% of cold ethanol for 2 min, respectively, a 10-µL probe solution was used for each slide, and then denaturing was performed for 2 min at 72°C–73°C. The hybridization was performed in a humid chamber for at least 16 h at 37°C. After hybridization, the slides were washed in 0.3% IGEPAL CA-630, Sigma-Aldrich (or NP-40)/0.4XSSC for 2 min at 73°C and then washed in 0.1% IGEPAL CA-630 Sigma-Aldrich (or NP-40)/2XSSC for 90 sec at room temperature. Finally, slides were counterstained with DAPI (4′,6-diamidino-2-phenylindole) solution (0.24 μg/mL, Sigma-Aldrich), and the images were captured with an Olympus BX-51 microscope equipped with a DP-70 CCD camera.

Aerenchyma formation was examined using the same roots that were used for the ROL measurements described above. Following determination of ROL, the root segments of 10–100 mm from the tips were stored in 50% ethanol for later anatomical observations. Segments of roots that had grown aerobically were taken after transferring to stagnant, deoxygenated conditions on day 0 (i.e. control) and on days 1, 2, 3, and 4. The segments were 4 mm long and were taken at 10-mm intervals with their centers at 20–100 mm from the tip. Transverse sections were made by hand using a razor blade, and were imaged using a BX60 light microscope with a DP70 CCD camera (both Olympus). The area occupied by aerenchyma was measured using the Image J software (version 1.46r; https://imagej.nih.gov/ij/) and is expressed as a percentage of the entire cross-sectional area of the root.

SDF-1 staining of human IVDs (nucleus pulposus and cartilaginous endplate, respectively) and the rat motion segment was quantified using a microscope and Image-Pro Plus ver. 6.0 software (Media Cybernetics, Silver Spring, MD, USA), according to the method developed by Xavie et al.12 (link) Briefly, an area of interest in each section was first selected at 40× magnification, and then 10 digital images at 1360 × 1024 pixel resolution and 400× magnification were captured with a DP 70 CCD camera (Olympus, Tokyo, Japan) coupled with an AX-70 microscope (Olympus). The measurement parameter was integrated optical density (IOD). Optical density was calibrated and the area of interest was set as follows: hue, 0-30; saturation, 0-255; intensity, 0-255. Then the values were counted.
The number of CXCR4 immunoreactive cells in human and rat specimens was expressed as a percentage of the total number of cells, which were randomly counted in 10 fields at ×400 magnification.