An effective histological staining process to visualize bone interstitial fluid space using confocal microscopy.
Bone is a composite porous material with two functional levels of porosity: the vascular porosity that surrounds blood vessels and the lacunar-canalicular porosity that surrounds the osteocytes. Both the vascular porosity and lacunar-canalicular porosity are directly involved in interstitial fluid flow, thought to play an important role in bone's maintenance. Because of the small dimensions of the lacunar-canalicular porosity, interstitial fluid space has been difficult to visualize and quantify. We report a new staining protocol that is reliable and easily reproducible, using fluorescein isothiocyanate (FITC) as a probe visualized by confocal microscopy. Reconstructed FITC-stained cross sections enable effective visualization of bone microstructure and microporosities. This new staining process can be used to analyze interstitial fluid space, providing high-resolution quantification of the vascular pores and the lacunar-canalicular network of cortical and cancellous bone.
Department of Biomedical Engineering, City College of New York, New York, NY 10031, USA.
Cesare
C
Ciani
Stephen B
SB
Doty
Susannah P
SP
Fritton
D000818:Animals; D045604:Extracellular Fluid / Q000378:metabolism*; D008297:Male; D018613:Microscopy, Confocal / Q000379:methods*; D010011:Osteocytes / Q000166:cytology* / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley