The histology and histomorphometry measurements were performed as previously described 19 (link). In brief, the right femurs were fixed in 10% Millonig's formalin with 0.5% sucrose for 24 h and were gradually dehydrated into 100% ethanol. The femurs were embedded undecalcified in methyl methacrylate and stained for Goldner trichrome staining. The histomorphometric examination of trabecular bone, osteoblast number, and osteoid volume, was done on 5 μm longitudinal sections with a digitizer tablet (OsteoMetrics, Inc., Decatur, GA, USA) interfaced to a Zeiss Axioscope (Carl Zeiss, Thornwood, NY, USA) with a drawing tube attachment. The right tibia was fixed in 10% Millonig's formalin for 24 h and were decalcified in 14% EDTA for 7-10 days. The bones were embedded in paraffin before obtaining 5-μm longitudinal sections. After removal of paraffin and rehydration, sections were stained for TRAP activity and counter-stained with haematoxylin and osteoclasts were enumerated on the trabecular bone surface.
Digitizer tablet
Manufactured by OsteoMetrics
Sourced in United States
The Digitizer tablet is a lab equipment device that digitizes handwritten or drawn input into a computer-readable format. It captures the position and movement of a stylus or other pointing device on its surface and converts it into digital data that can be processed and stored electronically.
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Lab products found in correlation
4 protocols using digitizer tablet
1
Histology and Histomorphometric Analysis of Bone
2
Bone Formation and Resorption Measurements
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The left femurs were embedded undecalcified in methyl methacrylate. The dynamic histomorphometric examination of trabecular bone formation was done on 5 μm longitudinal sections with a digitizer tablet (OsteoMetrics, Inc, Decatur, GA, USA) interfaced to a Zeiss Axioscope (Carl Zeiss, Thornwood, NY, USA) with a drawing tube attachment. The right femurs fixed and decalcified in 14% EDTA for 7–10 days. The bones were embedded in paraffin before obtaining 5 μm longitudinal sections. After removal of paraffin and rehydration, sections were stained for TRAP activity and counter-stained with fast green, and osteoclasts were enumerated on the trabecular bone surface using Osteomeasure histomorphometric software (OsteoMetrics, Inc, Decatur, GA, USA).
3
Bone Density and Microarchitecture Analysis in Mice
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Bone mineral density (BMD) was measured in live mice by dual-energy x-ray absorptiometry with a PIXImus Mouse Densitometer (GE Lunar Corp., Madison, WI) using the manufacturer’s software as described previously18 (link). Growth curves were obtained by sequential BMD measurement of the same animals from 8 weeks to 24 weeks of age. Micro-CT analysis of cortical and trabecular architecture was performed in femurs and fourth lumbar vertebrae (L4), as previously described19 (link). Biomechanical properties of femurs and L4 vertebrae were measured by 3-point bending and compression testing respectively, as previously described20 (link). L1–L3 lumbar vertebrae were fixed, embedded undecalcified in methylmethacrylate, and histomorphometric examination was done on longitudinal sections with a digitizer tablet (OsteoMetrics, Inc., Decatur, GA) interfaced to a Zeiss Axioscope (Carl Zeiss, Thornwood, NY) with a drawing tube attachment, as previously described21 (link). Terminology recommended by the Histomorphometry Nomenclature Committee of the American Society for Bone and Mineral Research was used in this study22 (link).
4
Comprehensive Bone Architecture and Biomechanics Analysis
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