1. Adapting Image J to Bone Histomorphometry
Kevin Egan

2. “Practitioners of Bone Histomorphometry communicate with each other in a variety of arcane languages, which in general are unintelligible to those outside the field.”
Parfitt et al. "Journal of Bone and Mineral Research 2(1987):

3. Background
Can be performed with either advanced imaging (microCT) or stained tissue sections
MicroCT provides more accurate measurement of the 3D structures.
But is limited to measuring mineralized surfaces and is expensive.
Stained sections allow for measuring of actively mineralizing surfaces.

4. Background
Quantitative analysis of stained tissue sections. Digital images of sections are taken and software measures basic defined parameters within the sample
Commercially available solutions require purchasing entire computer setups in additions to expensive proprietary software

5. Background
With a normal microscope and camera setup, researchers should be able to perform calculations without expensive software suites
Using Photoshop to automatically select the bone areas from an H/E stain, we can use Image J to measure the bone within the sample.

6. Image J
Image J counts the number of black pixels in the image. From these, it derives basic measurements.
Before working with complex samples, we need to make sure it accurately calculates these measurements
Distance between points
Area
Length (perimeter)
Number

7. Testing Create basic objects with easy to calculate dimensions.
2 * 2 inch square. Area: 4 inches
Perimeter: 8 inches
Create increasingly complex objects inside and see how they affect the calculations

9. βox within a box test

10. Circle within a circle test

11. Next steps Make sure Image J calculates perimeters correctly
Error is introduced when trying to measure circular objects which are composed of square pixels.
Circle with Diameter of 1 inch should have perimeter of 3.14 (2 * pi * r = 3.14), but image J calculates the perimeter to be 3.3
Will search for plugins which utilize different algorithms for calculating circular objects.