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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
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)
LengthArea
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
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%
Image J vs Manual calculation of Perimeter
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Diameter of circle (mm)
Per
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er (
mm
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Image J
manual
Difference between numbers vs % error
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Diameter of Circle (mm)
Difference betw eenvalues
% error
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%
Original vs adjusted % error
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Diameter of Circle (mm)
% e
rro
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Original % error
Variable Adjusted %error
Average adjusted %error
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