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Adapting Image J to perform Bone Histomorphometry

Adapting Image J to perform Bone Histomorphometry. Goal: Develop method by which Bone histomorphometry can be performed without cost by use of free software from the NIH. Bone histomorphometry uses stained bone slides to measure various bone parameters

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Adapting Image J to perform Bone Histomorphometry

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  1. Adapting Image J to perform Bone Histomorphometry Goal: Develop method by which Bone histomorphometry can be performed without cost by use of free software from the NIH.

  2. Bone histomorphometry uses stained bone slides to measure various bone parameters • Commercial software is used to perform the measurements • But these are all based on basic measurements of length, area, and number • Software specifically tailored towards bone field simplifies the measurements, but little competition leads to high prices • Image J can perform the basic same measurements (Area, length and number) Length Area

  3. Image J calculates area accurately • Compared Image J calculated vs manually calculated measurements of area from various samples. • Control sample was 2 * 2 square inch box • Started creating void areas within the box to make sure that image J can measure complicated images

  4. Image J introduces error with distance measurements of circles • On a computer screen, circles are made of square pixels • Comparing to manual calculations, image J introduced an average error of 5.4%

  5. Correcting for error introduced • Error was consistent in that Image J was over estimating the distance and a little over 5% • Excel equation can automatically remove the error introduced by Image J. • Average error was reduced from 5.4% to .29%

  6. Next steps • All bone histomorphometry is performed on undecalcified bone samples embedded in methyl methacrylate • We have some bone samples, we need to get the materials to do the embedding • Also need to figure out proper cutting angles for the polycut microtome • Analysis is performed on samples stained using standard H/E and Goldners Trichrome • H/E allows for measurement of bone and tissue volumes • Goldner trichrome reveals unmineralized bone matrix, allowing measurements of osteoid volumes

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