Semiautomated methods for cancellous bone histomorphometry using a general-purpose video image analysis system

J Microsc. 1994 Jan;173(Pt 1):53-66. doi: 10.1111/j.1365-2818.1994.tb03427.x.


Semiautomated methods are used to measure elongated, curved and complex branching profiles and isolated perimeter segments in monochrome video images with a general-purpose analysis system. These methods are used to make the major primary measurements of bone histomorphometry. Accuracy and reproducibility of the image acquisition, processing and measurement system is documented by measuring a semicircular standard of known dimensions. Semiautomated applications of the Ar/Le method for measuring areas and perimeters, and calculating lengths and widths of osteoid seams, lengths of mineralization labels and mineral apposition rate, wall width, indirect measurements of eroded, osteoclastic and osteoblastic perimeters without tracing, and measurement of mineralized or total cancellous bone area and perimeter gave values comparable to measurements of the same parameters by tracing or grid counting techniques with equal or better reproducibility and much greater efficiency. Intraindividual variation in measuring multiple bone biopsies methods. Major sources of variability for semi-automated methods were image magnification and selection of profile edges by thresholding, and sources of variability for manual methods are image magnification, numbers of orthogonal intercepts, tracing speed and accuracy of the algorithm used to measure traced pixels. Semiautomated methods are accurate, reproducible and rapid methods suitable for bone histomorphometry.

MeSH terms

  • Bone Remodeling
  • Bone and Bones / anatomy & histology*
  • Bone and Bones / physiology
  • Calcification, Physiologic
  • Humans
  • Image Processing, Computer-Assisted / methods*
  • Microscopy, Fluorescence / methods
  • Staining and Labeling