To begin analysis of the Hematoxylin and Eosin or HE images of the mouse bone tissue, open the HE images with case viewer software. Select the region of interest or ROI on the slide and save it as a color image. Then open the image in image J.Select the wand tool from the toolbar, and click on the trabecular bone.
To adjust the tolerance and contiguous settings as needed, go to image, then adjustments, followed by black and white. And click okay. Repeat the steps for other areas of the bone.
Invert the selection and fill it with white color. Sequentially click on process binary and convert to mask to convert the color image to a binary image. Use a plugin in the software to analyze structural parameters.
Run area by volume fraction to calculate bone volume to total volume of bone. The histopathological analysis performed by HE staining of mice femur showed the trabecular bones under the growth plate of the distal femur were reduced in the ovariectomized osteoporosis model mice when compared to the sham operated group. The quantitative analysis of tissue sections also indicated that the OVX mice had less trabecular bone area than those in the sham group.
