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Outline objects

This detection algorithm is based on the current image adjustments histogram, not the raw image data.

Procedure

1. Open an image (you can't detect on a live image) and zoom to an area containing objects representative of the objects you wish to outline.
2. Click Trace>Automatic>Outline objects.
3. In the Outline Detected Objects window, select Use Full Image or Use Region(s) of Interest under Detection Space.
   Use Region(s) of Interest

   • Click Use Existing ROIs to use existing contours.
     • Click a contour to select it; hold down Shift and click on contours to select multiple contours.
     • Click Done Selecting when satisfied with your selection.
   • Click Draw ROIs to trace a contour.
     • Trace a contour in the Tracing window.
     • Click Finished ROIs.
4. Click a representative object. Tissue Mapper displays a preview of outlined detected objects.
   • For a list of contour measurements, click Contour Measurements. Tissue Mapper displays the results in the Contour Measurements window.
   • If the outline is difficult to distinguish, change its color: Under Status / Change Preview Color, click to change the color of the outline.
   • To use presets : See Automatic Object Detection/Presets.
5. Adjust any of the parameters; every time you adjust a parameter, Tissue Mapper updates the Results .
   Parameters

   • Sensitivity: The higher the sensitivity, the smaller the detected object around the representative sample color (displayed under Object Color ).
     • Click Apply to refresh the preview.
     • To sample a different color, click on another color in the tissue.
   • Outline Point Density: The higher the density , the more precise the outline. Click Apply to refresh the preview.
   • Exclude objects: Excludes objects from detection that are Smaller than/ Larger than the set micron value; use in conjunction with sensitivity.
   • Separate Objects By Average Size: Use to increase the accuracy for the count of detected objects.

   Example: You define the average size as 10 µm2 and Tissue Mapper detects an object approximately 30 µm2.
   The program counts the object as 3 objects instead of 1.

   • Screen Edge Restrictions:Tissue Mapper ignores objects that meet the criteria.
6. Click Outline Objects to apply the parameters.