Undecalcified Bone Histomorphometry

Processing of undecalcified bone specimens and cancellous bone histomorphometry were performed as described^{25 (link),70 (link)}. Bone specimens (femur, tibia, and vertebra) were dehydrated and embedded in methylmethacrylate. Sections (5 μm in thickness) were prepared using a microtome (#RM2235, Leica) microtome and were stained by the von Kossa/nuclear fast red method. For hematoxylin and eosin (HE) and tartrate-resistant acid phosphatase (TRAP) staining, bone specimens were decalcified in 10% EDTA for 2 weeks at room temperature. After serial dehydration in a series of ethanol (70–100%), samples were embedded in paraffin. Sections (5 μm in thickness) were processed by a microtome (#RM2235, Leica) ready for staining. Histomorphometric measurements were performed using OsteoMeasure software (OsteoMetrics, Decatur, GA)^{71 (link)}.

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Wang Y., Deng P., Liu Y., Wu Y., Chen Y., Guo Y., Zhang S., Zheng X., Zhou L., Liu W., Li Q., Lin W., Qi X., Ou G., Wang C, & Yuan Q. (2020). Alpha-ketoglutarate ameliorates age-related osteoporosis via regulating histone methylations. Nature Communications, 11, 5596.

Publication 2020

RM2235 OsteoMeasure software

Bone Cancellous bone Dehydration Edta Embedded paraffin Eosin Ethanol Femur Hematoxylin Methylmethacrylate Microtome Tartrate resistant acid phosphatase Tibia Vertebra

Corresponding Organization :

Other organizations : Sichuan University, University of California, Los Angeles

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Protocol cited in 2 other protocols

Variable analysis

independent variables

Processing of undecalcified bone specimens
Cancellous bone histomorphometry

dependent variables

Histomorphometric measurements

control variables

Dehydration and embedding in methylmethacrylate
Staining methods (von Kossa/nuclear fast red, hematoxylin and eosin, tartrate-resistant acid phosphatase)
Microtome used for sectioning (Leica RM2235)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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