Microscope digital camera dp21

The collected epididymal WAT was fixed in formalin solution, dehydrated, paraffin-embedded, and sectioned. Sections (10 µm thick) were stained with hematoxylin and eosin, then examined and photographed under Olympus Microscope Digital Camera DP21 (Olympus China, LTD., Beijing, China). Adipocyte sizes were measured for at least 200 individual cells per mice with Image J software (National Institutes of Health, Bethesda, MD, USA).

Histomorphometry was performed to quantify the amount of new bone formed within the defect region. One picture was taken for each slice using a high-resolution camera (Microscope Digital Camera DP21; Olympus Corp.) mounted on the microscope at 4× magnification. Each picture included the whole defect, which was bordered by the upper and lower bony margins of the defect, the sheet covering the defect, and the lingual periosteum. All pictures were saved to a computer in TIFF format. They were analyzed using an ImageJ plugin distributed as a part of the Fiji project (https://imagej.net/Fiji) [19 (link),20 (link)]. The analyzing process was similar to that in the 2018 study by Gavazzoni, Filho, and Hernanes [21 (link)]: The total area as mentioned above (i.e., the whole defect) and the new bone area were selected using the selection tool in Fiji. Then, these selections were stored using the ROI manager tool. After that, the total areas were measured, and the percentage of new bone was calculated (Figure 2). This analysis was performed on all pictures taken from the rats sacrificed at weeks 4 and 8, because bone formation in both groups at week 2 was minimal or non-existent.

Histomorphometry was conducted to quantify the percentage of new bone formed within the defect region in a slide of each sample. One image at 1.25× magnification was taken for each slide using a high-resolution camera (Microscope Digital Camera DP21; Olympus Corp, Tokyo, Japan) mounted on the microscope. Each image included the whole defect region, the upper and lower bony margins of the defect region, the sheet covering the defect, and the lingual periosteum. Images were analyzed using a plugin in Fiji software [20 (link)]. The analysis method was similar to the approach used in a previous study [21 (link)]. Briefly, the total area as mentioned above (i.e., the whole defect) and the new bone area were selected using the selection tool in Fiji. These selections were saved using the region of interest (ROI) manager tool. Then, all areas were measured and the percentage of new bone was calculated using the formula: P = N/T × 100%, where P is the percentage of new bone area, N is the area of new bone area, and T is the area of the whole defect region (Figure 4).