http://imagej.273.s1.nabble.com/Disector-counting-frame-tp3688237p3688239.html
this an ingenious approach. I have some macro lines that prompt for user input regarding grid placement and will create the grid with a systematic random placement that I will send separately.
> I used this approach to produce an unbiased dissector frame, and it may help
> those searching under this topic. I recorded the process as a macro and ran
> it automatically once I had determined the sizes of the different squares.
> It can be run on individual images or on an image stack.
>
> Squares are defined in IJ by (X, Y, width, height), where XY is the
> right-hand top vertex. A random number generator and a spreadsheet can be
> used to calculate random XY co-ordinate start points, and so satisfy the
> requiremnts of an unbiased dissector.
>
> Set on Record Macro to capture procedures.
>
> From your thresholded binary image, first sample a square around 2x the size
> of the largest particle you want to count. Save as Sample image.
>
> Set the Scale to global. Create and name a square frame image of the same
> size as the Sample square. Use New image & choose the required pixel size.
> Draw a slightly smaller square inside and fill it to give a border of black
> ~ 5-10 pixels. Save this border square as a binary Frame image.
>
> Within the Sample image, draw a Count square to a known size (e.g. 1 mm2),
> starting from 0, 0 of the Sample image. Run Analyse Particles, Include On
> edges, Show Masks. Save the Mask as a binary Mask1 image. Save all Mask
> image as binary images, or later Image Calculator functions may not work.
>
> Return to Sample image and use the Polygon tool to create an L-shaped ROI
> the inverse of the Count square, which shares 3 common vertices with the
> Count square on the inside part of the L. Using this L-ROI run Analyse
> Particles, Include On edges, Show Masks. Save Mask as Mask2 image. Then run
> Analyse Particles, exclude On Edges, Show Masks. Save Mask as Mask3.
> Subtract Mask 3 from Mask 2 to get only the particles intersecting the edges
> of the L-shaped ROI. Save as Mask 2. Add Mask2 to Mask1, Add Frame. Save as
> Mask2 and run Analyse Particles, exclude On Edges, Show Masks.
>
> This gives you an image or a stack where the top and right-hand particles
> touching the Count square are excluded, while those jutting out of the
> bottom and left-hand have been stitched onto edges and are counted. There
> can be problems if your particles are too large for the Sample square and
> are truncated when they hit the border resulting from the Frame.
>
> For stacks, I found that large stacks work best where the image stacks are
> all saved onto the Desktop, the use of Create New Window is used only when
> necessary, and intermediate image stacks are saved (hence all the Save
> steps).
>
> Teresa Wegrzyn
> Institute of Food, Nutrition and Human Health
> Massey University
> Palmerston North New Zealand.
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> View this message in context:
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