Image plus 2

For histomorphology, liver samples fixed in buffered PFA were dehydrated in ethanol crescent series, embedded in historesin (Leica, Wetzlar, Germany), and the sections (3 μm thickness) were stained with Toluidine blue and basic fuchsin. The slides were analyzed using an Olympus BX51 light microscope (Olympus, Ballerup, Denmark) equipped with a camera connected to a computer using Olympus DP2-BSW software (Version 2.2, Olympus, Ballerup, Denmark, 2008). For the quantification of liver vascularization, the total area occupied by the histological section and the total area occupied by the blood vessels (arteries and veins) were measured with the help of the image analysis software (Motic Image Plus 2.0, Motic China Group Co., Ltd., Hong Kong, China, 2006). The fractional area of the blood vessels was calculated through the ratio between the total area occupied by these vessels and the total area occupied by the histological section (six-eight non-contiguous fields/section/fish), multiplied by 100. The mean values for each fish were calculated and these values were used to determine the average of each experimental condition. The presence of histological alterations was also evaluated by a randomized blind method.

Four birds per treatment were slaughtered and the medial region of the ileum was sampled for histology analyses. Samples were rinsed with sterile saline, immersed in buffered formalin for 24 hours and rinsed in 70% ethanol. Slides were mounted with 5μm sections and stained with hematoxylin-eosin. Two slides with five to seven sections were mounted per bird. Photomicrographs of the duodenal mucosa were taken using a digital camera with 12.1 megapixels (Sony Inc.) connected to a light microscope, with 1.7 optical zoom and 10X-objective. The images were analyzed with Image J software [19 ]. Villus height, crypt depth and villus:crypt ratio were determined. Villus height was measured from its apex to the basal region, crypt depth was measured from the crypt basis to the region of transition between the villus and crypt. For each animal, 30 measurements were taken, with a total of 120 measures per treatment per parameter. Ratio was calculated between villus height and crypt depth.
Goblet cell counts were performed in sections stained with Periodic Acid-Schiff (PAS) using a analysis software (Motic Image Plus 2.0, Motic, China) in 2000μm-linear segments of 12 repetitions per treatment (three photomicrographs from each of four slides).

Contact angle analysis, commonly applied to assess polymers surface energy, provided valuable information on the overall polymer wettability. The data are dependent on surface modifications (i.e., chemical composition, roughness, heterogeneity), which provide an indication of possible abiotic or biotic deterioration activities. Static contact angle measurements were performed by dropping 2 μL of Milli-Q water on the vacuum-dried polymer samples. Images were successively acquired by a home-made system that allowed translating the sample along the vertical and horizontal axis. Static contact angle was calculated by using the Motic Image Plus 2.0 software (Motic China Group Co.). The contact angles are the average of measurements carried out on three replicate analyses (three drops placed on three samples) at all the different immersion periods.

The measuring system presented in Figure 5 was developed to carry out the full scope of the study of the orthogonal cutting process of cortical bone tissue. The tool movement during orthogonal cutting was performed using UMT Bruker tribotester (Billerica, MA, USA) drives equipped with a 3-axis motion system and high-resolution stepper motors drives. The DFM-20 two-axis force sensor measured the force with a range of 0.05 to 235 N. The following features characterize it: measuring resolution of 0.01 N, non-linearity of 0.02% and sampling frequency of 1000 Hz. It provides precise measurement of force, position in three axes and AE generated during cutting.
The Motic optical microscope (Hongkong, China) with a microscope camera ensuring image registration was used to observe the cutting process. A camera with a maximum resolution of 2048 × 1536 pixels was used. Pictures were taken during chip formation. In addition, microscopic measurements of the chips were carried out on dedicated software (Motic Image Plus 2.0, Motic, Hongkong, China).