Application of reflected light microscopy to identify and quantitate resorption by isolated osteoclasts

J Bone Miner Res. 1991 Jul;6(7):661-71. doi: 10.1002/jbmr.5650060703.

Abstract

A simple technique has been developed to identify the resorption lacunae excavated by avian osteoclasts in vitro. Briefly, devitalized bovine bone wafers, with cells in situ, are fixed, stained with toluidine blue, and then examined by reflected light microscopy. Resorption pits are clearly visible using a x 10 objective, even when the resorbed areas are covered by a confluent layer of cells. The technique can be used to quantify the plan area, depth, and volume of the excavations. Depth and volume are assessed using the x 50 objective lens. This technique has been used to investigate the effects of retinyl acetate and EHDP on bone resorption.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bone Resorption / physiopathology*
  • Cattle
  • Chick Embryo
  • Diterpenes
  • Etidronic Acid / pharmacology
  • Histological Techniques
  • In Vitro Techniques
  • Microscopy / methods*
  • Osteoclasts / drug effects
  • Osteoclasts / physiology*
  • Retinyl Esters
  • Vitamin A / analogs & derivatives
  • Vitamin A / pharmacology

Substances

  • Diterpenes
  • Retinyl Esters
  • Vitamin A
  • retinol acetate
  • Etidronic Acid