Differentiation of cancellous bone and medullary bone in laying hens: a novel technique for image analysis

A selective staining technique for the identification and differentiation of cancellous bone from medullary bone of the laying hen by image analysis is described. Undecalcified Polymaster resin sections were oxidized in acidified potassium permanganate and oxalic acid before being immersed in an ammoniacal silver solution. The sections were reduced in formalin, fixed in sodium thiosulfate and counterstained in naphthalene black 10B which was dissolved in picric and acetic acids. Intensely stained cancellous bone was prominent with this technique compared with a paler medullary bone component which permitted the former to be easily recognized and measured by image analysis.