Quantifying Marrow Bone in Femur Sections

The paraformaldehyde fixed femur samples were decalcified (6 weeks; in 10% EDTA phosphate buffer solution), dehydrated (in ethyl alcohol), hyalinized (in xylene), and the proximal diaphysis was embedded in paraffin wax. Sample sections (5 μm thick; mounted on glass slides) were stained with toluidine blue and scanned using an optical microscope (BX46; Olympus, Japan). Three photomicrographs (40× magnification) were taken from each sample. The area percentage of MB is expressed as the percentage of MB in the region of the marrow cavity where the MB is located (ImageJ 1.8.0. software).

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Yan J., Wang J., Chen J., Shi H., Liao X., Pan C., Liu Y., Yang X., Ren Z, & Yang X. (2023). Adjusting phosphate feeding regimen according to daily rhythm increases eggshell quality via enhancing medullary bone remodeling in laying hens. Journal of Animal Science and Biotechnology, 14, 17.

Publication 2023

Buffer Cavity Diaphysis Edta Ethyl alcohol Femur Marrow Optical microscope Paraffin wax Paraformaldehyde Phosphate Photomicrographs Toluidine blue Xylene

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Other organizations : Northwest A&F University

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Variable analysis

independent variables

Duration of decalcification (6 weeks)

dependent variables

Area percentage of mineralized bone (MB) in the region of the marrow cavity where the MB is located

control variables

Paraformaldehyde fixed femur samples
10% EDTA phosphate buffer solution for decalcification
Dehydration in ethyl alcohol
Hyalinization in xylene
Embedding of the proximal diaphysis in paraffin wax
Sample sections of 5 μm thickness
Mounting of sample sections on glass slides
Staining with toluidine blue
Scanning using an optical microscope (BX46; Olympus, Japan)
Magnification of 40×

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