A two-color acid-free cartilage and bone stain for zebrafish larvae

Biotech Histochem. 2007 Feb;82(1):23-8. doi: 10.1080/10520290701333558.


Traditionally, cartilage is stained by alcian blue using acidic conditions to differentiate tissue staining. The acidic conditions are problematic when one wishes to stain the same specimen for mineralized bone with alizarin red, because acid demineralizes bone, which negatively affects bone staining. We have developed an acid-free method to stain cartilage and bone simultaneously in zebrafish larvae. This method has the additional advantage that PCR genotyping of stained specimens is possible.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Acids
  • Animals
  • Bone and Bones / anatomy & histology*
  • Cartilage / anatomy & histology*
  • Coloring Agents*
  • Histocytochemistry / methods*
  • Larva / anatomy & histology*
  • Magnesium Chloride*
  • Polymerase Chain Reaction
  • Zebrafish


  • Acids
  • Coloring Agents
  • Magnesium Chloride