Tetrachrome counter stain

Manufactured by Polysciences

Sourced in United States

Tetrachrome counter stain is a laboratory reagent used in microscopy and histology. It is a mixture of dyes that stains various cellular components, allowing for the differentiation and visualization of different structures within a biological sample.

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Lab products found in correlation

Osteomeasure system, by OsteoMetrics (1 mentions) Osteomeasure image analysis system, by OsteoMetrics (1 mentions)

2 protocols using tetrachrome counter stain

Histomorphometric Analysis of Vertebrae

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The histological methods used here have been previously described in detail [47] (link). In brief, 5^th lumbar vertebra were dehydrated in graded increases of ethanol and xylene and embedded in methyl methacrylate. Sections (4-µm thick) were cut with a vertical bed microtome (Leica/Jung 2165) and were stained according to the Von Kossa method with a tetrachrome counter stain (Polysciences, Warrington, PA, USA) for assessment of cell-based measurements. Osteoblast and osteoclast number and perimeter were measured in the entire cancellous compartment of the vertebral body. Osteoblast and osteoclast number was expressed per bone perimeter (#/mm) and osteoblast and osteoclast perimeter was expressed per total bone perimeter (%), bone area (mm/mm²) and tissue area (mm/mm²). Osteocyte number was also determined in the entire cancellous compartment of the vertebral body and expressed per total bone area (#/mm). Histomorphometric data were collected using the OsteoMeasure System (OsteoMetrics, Inc., Atlanta, GA, USA). All histomorphometric data are reported using standard 2-dimentional nomenclature [48] .

Gingery A., Subramaniam M., Pitel K.S., Reese J.M., Cicek M., Lindenmaier L.B., Ingle J.N., Goetz M.P., Turner R.T., Iwaniec U.T., Spelsberg T.C, & Hawse J.R. (2014). The Effects of a Novel Hormonal Breast Cancer Therapy, Endoxifen, on the Mouse Skeleton. PLoS ONE, 9(5), e98219.

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Histomorphometric Analysis of Bone Mineralization

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At 4 days and 1 day prior to sacrifice, mice were injected subcutaneously with calcein (10 mg/kg) to label mineralizing bone. Distal femora were processed for static and dynamic histomorphometry as previously described. (18 (link)) Sections were stained according to the Von Kossa method with a tetrachrome counter stain (Polysciences, Warrington, PA) for assessment of cell-based measurements. Unstained sections were used for assessment of bone area and dynamic histomorphometry. Bone area/tissue area (%), trabecular number (mm^-1), trabecular thickness (μm), trabecular spacing (μm), mineralizing perimeter/bone perimeter (double-labeled perimeter + ½ single-labeled perimeter, %), mineral apposition rate (μm/day), bone formation rate/bone perimeter (μm²/μm/y), bone formation rate/bone area (%/year) and osteoclast perimeter/bone perimeter (%) were determined using the Osteomeasure image analysis system (OsteoMetrics Inc., Atlanta, GA) as described. (18 (link)) Additionally, mineralizing perimeter/bone perimeter (%), mineral apposition rate (μm/day) and bone formation rate/bone perimeter (μm²/μm/y) were determined on the endocortex of the femoral diaphysis. All data are reported using standardized units and nomenclature for bone histomorphometry. (27 (link))

Hawse J.R., Pitel K.S., Cicek M., Philbrick K.A., Gingery A., Peters K.D., Syed F.A., Ingle J.N., Suman V.J., Iwaniec U.T., Turner R.T., Spelsberg T.C, & Subramaniam M. (2014). TGFβ Inducible Early Gene-1 Plays an Important Role in Mediating Estrogen Signaling in the Skeleton. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, 29(5), 1206-1216.

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