Powershot g7 x mark 2

For analysis of colony morphology, 1 µl of overnight cultures of the S. Enteritidis strains was used to spot onto LB agar plates without salt, containing 40 µg ml⁻¹ of Congo red (Sigma) and 20 µg ml⁻¹ of Coomassie brilliant blue (Sigma) (CR agar plates) or 200 µg ml⁻¹ of calcofluor white (Sigma) (CFW agar plates). The resulting colonies were grown at 26 °C for 96 h and visualized using a Stemi 305 stereomicroscope (Zeiss) equipped with an Axiocam 105 colour camera (Zeiss) or photographed using a digital camera (PowerShot G7X Mark II; Canon).

Our study protocol is shown in Figure 1. Eligible patients were identified in both inpatient and outpatient settings. Baseline demographics and clinical profile were collected prior to the commencement of the study. The approximate study duration for each patient is five visits or until complete resolution of the ulcer, whichever was earlier. All patients who were included in the study were subjected to a standardised DFU management pathway with standardised follow‐up; no additional clinic visits were required for the purpose of this study. During each clinic visit, wound measurements were recorded traditionally by a trained specialised wound nurse and electronically by a dedicated research coordinator using the C4W imaging system. An additional digital wound image capture with a reference ruler was also taken independently with a Canon (PowerShot G7X Mark II) digital camera. A wound episode was defined as any clinic consultation for this study. The total number of wound images is calculated by the number of images taken by each of the C4W imaging system (ie, each wound episode should result in nine wound images, with three wound images taken per device).

Clone formation assay was used to detect the cell proliferative ability. The transfected cells were seeded into a six-well cell culture plate with the same cell density (500 cells per well). The cells were preserved in a CO₂ incubator for 14 days, and the medium was replaced every 3 days. The cells were then rinsed with PBS, fixed with methanol for 20 min and stained with 0.1% crystal violet (cat. no. C0121-500ml, Beyotime Institute of Biotechnology) for 10 min at room temperature, and photographed using a digital camera (PowerShot G7 X Mark II, Canon). The average area of cell clusters in the images was analyzed using ImageJ software (version 1.45s/Java1.6.0_20, National Institutes of Health). All data are presented as the mean ± SD of five independent experiments.

The cerebral infarct volume and integrity of the BBB were assessed by TTC and Evans blue staining, respectively, as previously described (38 (link)). Briefly, Evans blue dye (2%; 4 ml/kg) was injected into the left femoral vein and allowed to circulate for 60 sec before the rats were euthanized with 1% pentobarbital (40 mg/kg) and perfused transcardially with PBS for 60 sec at 37°C. The brains were removed, washed with PBS and kept at −20°C for 30 min before sectioning. Coronal sections (2 mm) were cut in a rat brain matrix and stained with 2% TTC (Amresco, LLC) for 30 min at 37°C. Following staining, the brain slices were fixed with 4% paraformaldehyde for 10 min at 37°C to conserve the area stained by TTC. The stained brain sections were digitally photographed using a digital camera (PowerShot G7 X Mark II Canon, Inc.). The infarcted area (white) and BBB disruption (blue) of each brain section was measured using Image-Pro Plus 6.0 software (Media Cybernetics, Inc.). The infarcted volume and the BBB disrupted area were calculated according to the formula: 100% × (ipsilateral volume-contralateral volume)/contralateral volume.