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@lgelman

lgelman/Find_Wells_in_Plate

Last active August 29, 2015 14:11
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Finding wells in a multi-well plate
Raw
Find_Wells_in_Plate
// ******************************************************************************
// Initialisation: Close all windows (images, results, etc)
// ******************************************************************************
run("Set Measurements...", "center redirect=None decimal=9");
// Close all windows
run("Close All");
// Clears Log Window
print("\\Clear");
// Close Result window
if (isOpen("Results")==1) {
selectWindow("Results");
run("Close");
}
// Close Summary window
if (isOpen("Summary")==1) {
selectWindow("Summary");
run("Close");
}
// Clear ROI manager
count = roiManager("count");
if (count >= 1) {
allRegions = newArray(count);
roiManager("Select", allRegions);
roiManager("Delete");
}
print("Finding Well(s) border(s)... Please wait...");
rename("ORIG");
// ******************************************************************************
// Make a duplicate of original pictures and process it for convolution step.
// NB: Image is resized to speed up convolution process
// ******************************************************************************
selectWindow("ORIG");
run("Duplicate...", "title=FindCenters");
run("8-bit");
run("Invert");
run("Scale...", "x=0.04 y=0.04 width=757 height=592 interpolation=Bilinear average create title=FindCentersResized");
selectWindow("FindCenters");
close();
selectWindow("FindCentersResized");
run("32-bit");
run("Find Edges");
// ******************************************************************************
// Convolve the image with a kernel corresponding to a thick circle (= border of a well).
// NB: Kernel must be adapted to the size of the image
// ******************************************************************************
// Kernel 12.5x zoom on Axiozoom Z1 (Zeiss)
// Kernel is created from a binary image with a thick circle and save as a txt image.
// The thick circle matches the size of a well in the resized&processed original image (here Named FindCentersResized)
run("Convolve...", "text1=[0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 145 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 145 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0\n0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 26 77 77 77 77 77 77 77 77 178 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 178 77 77 77 77 77 77 77 77 26 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0\n0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 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255 255 254 255 255 255 255 255 254 255 255 232 132 141 17 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0\n0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 136 207 212 255 255 255 255 254 255 255 255 254 255 255 255 255 255 211 48 58 58 63 38 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 38 63 58 58 48 211 255 255 255 255 254 255 255 255 255 254 255 255 255 255 212 207 136 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0\n0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 52 252 245 242 249 255 255 255 255 255 255 255 255 255 254 255 255 255 255 255 186 0 12 12 12 12 10 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 12 12 12 12 12 12 0 186 255 255 255 255 255 254 255 255 255 255 255 255 255 255 255 249 242 245 252 52 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0\n0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 4 21 21 2 142 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 250 233 234 234 234 245 206 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 34 239 237 234 234 234 234 233 250 255 255 255 255 255 255 255 255 255 255 254 255 255 255 255 142 2 21 21 4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0\n0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 31 70 84 255 255 255 255 255 255 254 255 255 255 255 255 254 255 255 255 255 255 255 255 186 182 184 184 184 184 184 184 184 184 184 184 184 184 184 184 184 184 184 184 184 184 184 180 199 255 255 255 255 255 255 255 255 254 255 255 255 255 255 254 255 255 255 255 223 63 70 77 31 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0\n0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 123 137 132 128 143 255 255 255 255 254 255 255 255 255 255 255 255 255 254 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 143 128 141 95 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0\n0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 182 198 188 213 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 199 190 193 198 182 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0\n0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 63 253 241 240 240 240 242 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 253 240 240 240 240 247 230 13 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0\n0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 4 18 18 17 17 8 43 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 254 255 255 255 255 255 255 255 255 255 255 216 12 15 17 17 18 17 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0\n0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 11 73 77 77 77 77 77 77 77 77 108 251 255 255 255 255 255 255 255 255 255 255 255 255 255 216 77 77 77 77 77 77 77 77 77 58 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0\n]");
// ******************************************************************************
// Find centers of wells
//
// ******************************************************************************
// Crop image to remove artefacts at the border.
w = getWidth();
h = getHeight();
lar = 60;
makeRectangle(w/2-lar/2, h/2-lar/2, lar, lar);
setBackgroundColor(0, 0, 0);
run("Clear Outside");
// Keep only intense regions and find objects
getStatistics(a, m, mi, ma);
setThreshold(ma*0.7, ma);
setOption("BlackBackground", true);
run("Convert to Mask");
run("Open");
run("Analyze Particles...", " circularity=0.7-1.00 add");
selectWindow("FindCentersResized");
close();
count = roiManager("count");
print ("Number of Centers identified: "+count);
// ******************************************************************************
// Creates an array with all Well center coordinates
//
// ******************************************************************************
XCenter = newArray(count);
YCenter = newArray(count);
for (j=0; j<count; j++) {
roiManager("Select", j);
run("Measure");
XCenter[j] = 25*getResult("XM", j);
YCenter[j] = 25*getResult("YM", j);
}
// Delete regions from ROI manager
allRegions = newArray(count);
roiManager("Select", allRegions); roiManager("Delete");
// ******************************************************************************
// Draws circles correponding to wells in the original image
//
// ******************************************************************************
selectWindow("ORIG");
diameter = 5000;
for (k=0; k<count; k++) {
makeOval(XCenter[k]-diameter/2, YCenter[k]-diameter/2, diameter, diameter);
roiManager("Add");
}
roiManager("Show All");
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