PMID: 58925May 1, 1976

Light microscopic localization of labile calcium in hypertrophied chondrocytes of long bone with alizarin red S

The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
H K Kashiwa, H Z Park

Abstract

A method which localizes labile 5% ethylene glycol-bis-(beta-amino-ethyl ether)N-N'-tetraacetic acid-removable calcium in spherules within hypertrophied chondrocytes and in pericellular matrix using alizarin red S (ARS) is described. Fresh blocks of epiphyseal cartilage approximately 1 mm thick were immersed into 0.5-2% ARS solution containing 7% mounted on glass slides in 7% sucrose or in glycerol-gelatin. The stained tissue blocks were also dehydrated in acetone, cleared in xylene and mounted in Preservaslide. The ARS precipitated ionic calcium as red Ca-ARS salt which was birefringent in polarizing microscope, stable in water at pH 4-9 and in nonpolar organic solvent but soluble in polar solvents, especially in dimethyl sulfoxide. In contrast, ARS-stained insoluble calcium phosphate was stable even in dimethyl sulfoxide. Calcium in the hypertrophied chondrocytes, therefore, was thought to be present in a readily ionizable state instead of as insoluble calcium phosphate. Since addition of 7% sucrose retained as well as improved ARS localization of cellular calcium, the calcium was believed to be present in an osmotically sensitive, membrane-bound cytoplasmic compartment. The ARS-positive labile calcium in spherules which deve...Continue Reading

Citations

May 1, 1978·The Anatomical Record·K J Kuhar, D R Eisenmann
Apr 18, 2008·European Journal of Cell Biology·Zhidao XiaJ T Triffitt

Related Concepts

Birefringence
Skeletal Bone
Calcium
Cartilage
Chick Embryo
Cytochemistry
Hypertrophy
Staining and Labeling

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