Bone histomorphometry using free and commonly available software
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Updated: May 19, 2026

Automated Quantification of Hematopoietic Cell – Stromal Cell Interactions in Histological Images of Undecalcified Bone
Published on: April 8, 2015
Kevin P Egan1, Tracy A Brennan, Robert J Pignolo
1Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Aims:
Histomorphometric analysis is a widely used technique to assess changes in tissue structure and function. Commercially available programs that measure histomorphometric parameters can be cost-prohibitive. In this study, we compared an inexpensive method of histomorphometry to a current proprietary software program.
Methods And Results:
Image J and Adobe Photoshop(®) were used to measure static and kinetic bone histomorphometric parameters. Photomicrographs of Goldner's trichrome-stained femurs were used to generate black-and-white image masks, representing bone and non-bone tissue, respectively, in Adobe Photoshop(®) . The masks were used to quantify histomorphometric parameters (bone volume, tissue volume, osteoid volume, mineralizing surface and interlabel width) in Image J. The resultant values obtained using Image J and the proprietary software were compared and differences found to be statistically non-significant.
Conclusions:
The wide-ranging use of histomorphometric analysis for assessing the basic morphology of tissue components makes it important to have affordable and accurate measurement options available for a diverse range of applications. Here we have developed and validated an approach to histomorphometry using commonly and freely available software that is comparable to a much more costly, commercially available software program.

