void drawOutline(ImageProcessor ip, Roi roi, int count) {
if (showChoice == OVERLAY_OUTLINES || showChoice == OVERLAY_MASKS) {
if (overlay == null) {
overlay = new Overlay();
overlay.drawLabels(true);
overlay.setLabelFont(new Font("SansSerif", Font.PLAIN, fontSize));
}
Roi roi2 = (Roi) roi.clone();
roi2.setStrokeColor(Color.cyan);
if (lineWidth != 1) roi2.setStrokeWidth(lineWidth);
if (showChoice == OVERLAY_MASKS) roi2.setFillColor(Color.cyan);
overlay.add(roi2);
} else {
Rectangle r = roi.getBounds();
int nPoints = ((PolygonRoi) roi).getNCoordinates();
int[] xp = ((PolygonRoi) roi).getXCoordinates();
int[] yp = ((PolygonRoi) roi).getYCoordinates();
int x = r.x, y = r.y;
if (!inSituShow) ip.setValue(0.0);
ip.moveTo(x + xp[0], y + yp[0]);
for (int i = 1; i < nPoints; i++) ip.lineTo(x + xp[i], y + yp[i]);
ip.lineTo(x + xp[0], y + yp[0]);
if (showChoice != BARE_OUTLINES) {
String s = ResultsTable.d2s(count, 0);
ip.moveTo(r.x + r.width / 2 - ip.getStringWidth(s) / 2, r.y + r.height / 2 + fontSize / 2);
if (!inSituShow) ip.setValue(1.0);
ip.drawString(s);
}
}
}
void createMask(ImagePlus imp) {
Roi roi = imp.getRoi();
boolean useInvertingLut = Prefs.useInvertingLut;
Prefs.useInvertingLut = false;
if (roi == null || !(roi.isArea() || roi.getType() == Roi.POINT)) {
createMaskFromThreshold(imp);
Prefs.useInvertingLut = useInvertingLut;
return;
}
ImagePlus maskImp = null;
Frame frame = WindowManager.getFrame("Mask");
if (frame != null && (frame instanceof ImageWindow))
maskImp = ((ImageWindow) frame).getImagePlus();
if (maskImp == null) {
ImageProcessor ip = new ByteProcessor(imp.getWidth(), imp.getHeight());
if (!Prefs.blackBackground) ip.invertLut();
maskImp = new ImagePlus("Mask", ip);
maskImp.show();
}
ImageProcessor ip = maskImp.getProcessor();
ip.setRoi(roi);
ip.setValue(255);
ip.fill(ip.getMask());
maskImp.updateAndDraw();
Prefs.useInvertingLut = useInvertingLut;
}
boolean eraseOutsideRoi(ImageProcessor ip, Rectangle r, ImageProcessor mask) {
int width = ip.getWidth();
int height = ip.getHeight();
ip.setRoi(r);
if (excludeEdgeParticles && polygon != null) {
ImageStatistics stats = ImageStatistics.getStatistics(ip, MIN_MAX, null);
if (fillColor >= stats.min && fillColor <= stats.max) {
double replaceColor = level1 - 1.0;
if (replaceColor < 0.0 || replaceColor == fillColor) {
replaceColor = level2 + 1.0;
int maxColor = imageType == BYTE ? 255 : 65535;
if (replaceColor > maxColor || replaceColor == fillColor) {
IJ.error("Particle Analyzer", "Unable to remove edge particles");
return false;
}
}
for (int y = minY; y < maxY; y++) {
for (int x = minX; x < maxX; x++) {
int v = ip.getPixel(x, y);
if (v == fillColor) ip.putPixel(x, y, (int) replaceColor);
}
}
}
}
ip.setValue(fillColor);
if (mask != null) {
mask = mask.duplicate();
mask.invert();
ip.fill(mask);
}
ip.setRoi(0, 0, r.x, height);
ip.fill();
ip.setRoi(r.x, 0, r.width, r.y);
ip.fill();
ip.setRoi(r.x, r.y + r.height, r.width, height - (r.y + r.height));
ip.fill();
ip.setRoi(r.x + r.width, 0, width - (r.x + r.width), height);
ip.fill();
ip.resetRoi();
// IJ.log("erase: "+fillColor+" "+level1+" "+level2+" "+excludeEdgeParticles);
// (new ImagePlus("ip2", ip.duplicate())).show();
return true;
}
void createMask(ImagePlus imp) {
Roi roi = imp.getRoi();
boolean useInvertingLut = Prefs.useInvertingLut;
Prefs.useInvertingLut = false;
boolean selectAll =
roi != null
&& roi.getType() == Roi.RECTANGLE
&& roi.getBounds().width == imp.getWidth()
&& roi.getBounds().height == imp.getHeight()
&& imp.isThreshold();
if (roi == null || !(roi.isArea() || roi.getType() == Roi.POINT) || selectAll) {
createMaskFromThreshold(imp);
Prefs.useInvertingLut = useInvertingLut;
return;
}
ImagePlus maskImp = null;
Frame frame = WindowManager.getFrame("Mask");
if (frame != null && (frame instanceof ImageWindow))
maskImp = ((ImageWindow) frame).getImagePlus();
if (maskImp == null) {
ImageProcessor ip = new ByteProcessor(imp.getWidth(), imp.getHeight());
if (!Prefs.blackBackground) ip.invertLut();
maskImp = new ImagePlus("Mask", ip);
maskImp.show();
}
ImageProcessor ip = maskImp.getProcessor();
ip.setRoi(roi);
ip.setValue(255);
ip.fill(ip.getMask());
Calibration cal = imp.getCalibration();
if (cal.scaled()) {
Calibration cal2 = maskImp.getCalibration();
cal2.pixelWidth = cal.pixelWidth;
cal2.pixelHeight = cal.pixelHeight;
cal2.setUnit(cal.getUnit());
}
maskImp.updateAndRepaintWindow();
Prefs.useInvertingLut = useInvertingLut;
}
public void run(ImageProcessor ip) {
String[] imageNames = getOpenImageNames();
if (imageNames[0] == "None") {
IJ.error("need at least 2 binary open images");
return;
}
double previousMinOverlap = Prefs.get("BVTB.BinaryFeatureExtractor.minOverlap", 0);
boolean previousCombine = Prefs.get("BVTB.BinaryFeatureExtractor.combine", false);
GenericDialog gd = new GenericDialog("Binary Feature Extractor");
gd.addChoice("Objects image", imageNames, imageNames[0]);
gd.addChoice("Selector image", imageNames, imageNames[1]);
gd.addNumericField("Object_overlap in % (0=off)", previousMinOverlap, 0, 9, "");
gd.addCheckbox("Combine objects and selectors", previousCombine);
gd.addCheckbox("Count output", true);
gd.addCheckbox("Analysis tables", false);
gd.showDialog();
if (gd.wasCanceled()) {
return;
}
String objectsImgTitle = gd.getNextChoice();
String selectorsImgTitle = gd.getNextChoice();
double minOverlap = gd.getNextNumber();
boolean combineImages = gd.getNextBoolean();
boolean showCountOutput = gd.getNextBoolean();
boolean showAnalysis = gd.getNextBoolean();
if (gd.invalidNumber() || minOverlap < 0 || minOverlap > 100) {
IJ.error("invalid number");
return;
}
Prefs.set("BVTB.BinaryFeatureExtractor.minOverlap", minOverlap);
Prefs.set("BVTB.BinaryFeatureExtractor.combine", combineImages);
if (objectsImgTitle.equals(selectorsImgTitle)) {
IJ.error("images need to be different");
return;
}
ImagePlus objectsImp = WindowManager.getImage(objectsImgTitle);
ImageProcessor objectsIP = objectsImp.getProcessor();
ImagePlus selectorsImp = WindowManager.getImage(selectorsImgTitle);
ImageProcessor selectorsIP = selectorsImp.getProcessor();
if (!objectsIP.isBinary() || !selectorsIP.isBinary()) {
IJ.error("works with 8-bit binary images only");
return;
}
if ((objectsImp.getWidth() != selectorsImp.getWidth())
|| objectsImp.getHeight() != selectorsImp.getHeight()) {
IJ.error("images need to be of the same size");
return;
}
// close any existing RoiManager before instantiating a new one for this analysis
RoiManager oldRM = RoiManager.getInstance2();
if (oldRM != null) {
oldRM.close();
}
RoiManager objectsRM = new RoiManager(true);
ResultsTable objectsRT = new ResultsTable();
ParticleAnalyzer analyzeObjects =
new ParticleAnalyzer(analyzerOptions, measurementFlags, objectsRT, 0.0, 999999999.9);
analyzeObjects.setRoiManager(objectsRM);
analyzeObjects.analyze(objectsImp);
objectsRM.runCommand("Show None");
int objectNumber = objectsRT.getCounter();
Roi[] objectRoi = objectsRM.getRoisAsArray();
ResultsTable measureSelectorsRT = new ResultsTable();
Analyzer overlapAnalyzer = new Analyzer(selectorsImp, measurementFlags, measureSelectorsRT);
ImagePlus outputImp =
IJ.createImage("output", "8-bit black", objectsImp.getWidth(), objectsImp.getHeight(), 1);
ImageProcessor outputIP = outputImp.getProcessor();
double[] measuredOverlap = new double[objectNumber];
outputIP.setValue(255.0);
for (int o = 0; o < objectNumber; o++) {
selectorsImp.killRoi();
selectorsImp.setRoi(objectRoi[o]);
overlapAnalyzer.measure();
measuredOverlap[o] = measureSelectorsRT.getValue("%Area", o);
if (minOverlap != 0.0 && measuredOverlap[o] >= minOverlap) {
outputIP.fill(objectRoi[o]);
finalCount++;
} else if (minOverlap == 0.0 && measuredOverlap[o] > 0.0) {
outputIP.fill(objectRoi[o]);
finalCount++;
}
}
// measureSelectorsRT.show("Objects");
selectorsImp.killRoi();
RoiManager selectorRM = new RoiManager(true);
ResultsTable selectorRT = new ResultsTable();
ParticleAnalyzer.setRoiManager(selectorRM);
ParticleAnalyzer analyzeSelectors =
new ParticleAnalyzer(analyzerOptions, measurementFlags, selectorRT, 0.0, 999999999.9);
analyzeSelectors.analyze(selectorsImp);
selectorRM.runCommand("Show None");
int selectorNumber = selectorRT.getCounter();
if (combineImages) {
outputImp.updateAndDraw();
Roi[] selectorRoi = selectorRM.getRoisAsArray();
ResultsTable measureObjectsRT = new ResultsTable();
Analyzer selectorAnalyzer = new Analyzer(outputImp, measurementFlags, measureObjectsRT);
double[] selectorOverlap = new double[selectorNumber];
outputIP.setValue(255.0);
for (int s = 0; s < selectorNumber; s++) {
outputImp.killRoi();
outputImp.setRoi(selectorRoi[s]);
selectorAnalyzer.measure();
selectorOverlap[s] = measureObjectsRT.getValue("%Area", s);
if (selectorOverlap[s] > 0.0d) {
outputIP.fill(selectorRoi[s]);
}
}
selectorRoi = null;
selectorAnalyzer = null;
measureObjectsRT = null;
}
// selectorRT.show("Selectors");
outputImp.killRoi();
String outputImageTitle = WindowManager.getUniqueName("Extracted_" + objectsImgTitle);
outputImp.setTitle(outputImageTitle);
outputImp.show();
outputImp.changes = true;
if (showCountOutput) {
String[] openTextWindows = WindowManager.getNonImageTitles();
boolean makeNewTable = true;
for (int w = 0; w < openTextWindows.length; w++) {
if (openTextWindows[w].equals("BFE_Results")) {
makeNewTable = false;
}
}
TextWindow existingCountTable = ResultsTable.getResultsWindow();
if (makeNewTable) {
countTable = new ResultsTable();
countTable.setPrecision(0);
countTable.setValue("Image", 0, outputImageTitle);
countTable.setValue("Objects", 0, objectNumber);
countTable.setValue("Selectors", 0, selectorNumber);
countTable.setValue("Extracted", 0, finalCount);
countTable.show("BFE_Results");
} else {
IJ.renameResults("BFE_Results", "Results");
countTable = ResultsTable.getResultsTable();
countTable.setPrecision(0);
countTable.incrementCounter();
countTable.addValue("Image", outputImageTitle);
countTable.addValue("Objects", objectNumber);
countTable.addValue("Selectors", selectorNumber);
countTable.addValue("Extracted", finalCount);
IJ.renameResults("Results", "BFE_Results");
countTable.show("BFE_Results");
}
}
if (showAnalysis) {
ResultsTable extractedRT = new ResultsTable();
ParticleAnalyzer analyzeExtracted =
new ParticleAnalyzer(
ParticleAnalyzer.CLEAR_WORKSHEET | ParticleAnalyzer.RECORD_STARTS,
measurementFlags,
extractedRT,
0.0,
999999999.9);
analyzeExtracted.analyze(outputImp);
objectsRT.show("Objects");
selectorRT.show("Selectors");
extractedRT.show("Extracted");
} else {
objectsRT = null;
selectorRT = null;
}
objectsRM = null;
measureSelectorsRT = null;
analyzeObjects = null;
overlapAnalyzer = null;
objectRoi = null;
selectorRM = null;
objectsImp.killRoi();
objectsImp.changes = false;
selectorsImp.changes = false;
}
void drawRoiFilledParticle(ImageProcessor ip, Roi roi, ImageProcessor mask, int count) {
int grayLevel = (count < 65535) ? count : 65535;
ip.setValue((double) grayLevel);
ip.setRoi(roi.getBounds());
ip.fill(mask);
}
void analyzeParticle(int x, int y, ImagePlus imp, ImageProcessor ip) {
// Wand wand = new Wand(ip);
ImageProcessor ip2 = redirectIP != null ? redirectIP : ip;
wand.autoOutline(x, y, level1, level2, wandMode);
if (wand.npoints == 0) {
IJ.log("wand error: " + x + " " + y);
return;
}
Roi roi = new PolygonRoi(wand.xpoints, wand.ypoints, wand.npoints, roiType);
Rectangle r = roi.getBounds();
if (r.width > 1 && r.height > 1) {
PolygonRoi proi = (PolygonRoi) roi;
pf.setPolygon(proi.getXCoordinates(), proi.getYCoordinates(), proi.getNCoordinates());
ip2.setMask(pf.getMask(r.width, r.height));
if (floodFill) ff.particleAnalyzerFill(x, y, level1, level2, ip2.getMask(), r);
}
ip2.setRoi(r);
ip.setValue(fillColor);
ImageStatistics stats = getStatistics(ip2, measurements, calibration);
boolean include = true;
if (excludeEdgeParticles) {
if (r.x == minX || r.y == minY || r.x + r.width == maxX || r.y + r.height == maxY)
include = false;
if (polygon != null) {
Rectangle bounds = roi.getBounds();
int x1 = bounds.x + wand.xpoints[wand.npoints - 1];
int y1 = bounds.y + wand.ypoints[wand.npoints - 1];
int x2, y2;
for (int i = 0; i < wand.npoints; i++) {
x2 = bounds.x + wand.xpoints[i];
y2 = bounds.y + wand.ypoints[i];
if (!polygon.contains(x2, y2)) {
include = false;
break;
}
if ((x1 == x2 && ip.getPixel(x1, y1 - 1) == fillColor)
|| (y1 == y2 && ip.getPixel(x1 - 1, y1) == fillColor)) {
include = false;
break;
}
x1 = x2;
y1 = y2;
}
}
}
ImageProcessor mask = ip2.getMask();
if (minCircularity > 0.0 || maxCircularity < 1.0) {
double perimeter = roi.getLength();
double circularity =
perimeter == 0.0 ? 0.0 : 4.0 * Math.PI * (stats.pixelCount / (perimeter * perimeter));
if (circularity > 1.0) circularity = 1.0;
// IJ.log(circularity+" "+perimeter+" "+stats.area);
if (circularity < minCircularity || circularity > maxCircularity) include = false;
}
if (stats.pixelCount >= minSize && stats.pixelCount <= maxSize && include) {
particleCount++;
if (roiNeedsImage) roi.setImage(imp);
stats.xstart = x;
stats.ystart = y;
saveResults(stats, roi);
if (showChoice != NOTHING) drawParticle(drawIP, roi, stats, mask);
}
if (redirectIP != null) ip.setRoi(r);
ip.fill(mask);
}
/**
* Performs particle analysis on the specified ImagePlus and ImageProcessor. Returns false if
* there is an error.
*/
public boolean analyze(ImagePlus imp, ImageProcessor ip) {
if (this.imp == null) this.imp = imp;
showResults = (options & SHOW_RESULTS) != 0;
excludeEdgeParticles = (options & EXCLUDE_EDGE_PARTICLES) != 0;
resetCounter = (options & CLEAR_WORKSHEET) != 0;
showProgress = (options & SHOW_PROGRESS) != 0;
floodFill = (options & INCLUDE_HOLES) == 0;
recordStarts = (options & RECORD_STARTS) != 0;
addToManager = (options & ADD_TO_MANAGER) != 0;
displaySummary = (options & DISPLAY_SUMMARY) != 0;
inSituShow = (options & IN_SITU_SHOW) != 0;
outputImage = null;
ip.snapshot();
ip.setProgressBar(null);
if (Analyzer.isRedirectImage()) {
redirectImp = Analyzer.getRedirectImage(imp);
if (redirectImp == null) return false;
int depth = redirectImp.getStackSize();
if (depth > 1 && depth == imp.getStackSize()) {
ImageStack redirectStack = redirectImp.getStack();
redirectIP = redirectStack.getProcessor(imp.getCurrentSlice());
} else redirectIP = redirectImp.getProcessor();
} else if (imp.getType() == ImagePlus.COLOR_RGB) {
ImagePlus original = (ImagePlus) imp.getProperty("OriginalImage");
if (original != null
&& original.getWidth() == imp.getWidth()
&& original.getHeight() == imp.getHeight()) {
redirectImp = original;
redirectIP = original.getProcessor();
}
}
if (!setThresholdLevels(imp, ip)) return false;
width = ip.getWidth();
height = ip.getHeight();
if (!(showChoice == NOTHING || showChoice == OVERLAY_OUTLINES || showChoice == OVERLAY_MASKS)) {
blackBackground = Prefs.blackBackground && inSituShow;
if (slice == 1) outlines = new ImageStack(width, height);
if (showChoice == ROI_MASKS) drawIP = new ShortProcessor(width, height);
else drawIP = new ByteProcessor(width, height);
drawIP.setLineWidth(lineWidth);
if (showChoice == ROI_MASKS) {
} // Place holder for now...
else if (showChoice == MASKS && !blackBackground) drawIP.invertLut();
else if (showChoice == OUTLINES) {
if (!inSituShow) {
if (customLut == null) makeCustomLut();
drawIP.setColorModel(customLut);
}
drawIP.setFont(new Font("SansSerif", Font.PLAIN, fontSize));
if (fontSize > 12 && inSituShow) drawIP.setAntialiasedText(true);
}
outlines.addSlice(null, drawIP);
if (showChoice == ROI_MASKS || blackBackground) {
drawIP.setColor(Color.black);
drawIP.fill();
drawIP.setColor(Color.white);
} else {
drawIP.setColor(Color.white);
drawIP.fill();
drawIP.setColor(Color.black);
}
}
calibration = redirectImp != null ? redirectImp.getCalibration() : imp.getCalibration();
if (rt == null) {
rt = Analyzer.getResultsTable();
analyzer = new Analyzer(imp);
} else analyzer = new Analyzer(imp, measurements, rt);
if (resetCounter && slice == 1) {
if (!Analyzer.resetCounter()) return false;
}
beginningCount = Analyzer.getCounter();
byte[] pixels = null;
if (ip instanceof ByteProcessor) pixels = (byte[]) ip.getPixels();
if (r == null) {
r = ip.getRoi();
mask = ip.getMask();
if (displaySummary) {
if (mask != null) totalArea = ImageStatistics.getStatistics(ip, AREA, calibration).area;
else totalArea = r.width * calibration.pixelWidth * r.height * calibration.pixelHeight;
}
}
minX = r.x;
maxX = r.x + r.width;
minY = r.y;
maxY = r.y + r.height;
if (r.width < width || r.height < height || mask != null) {
if (!eraseOutsideRoi(ip, r, mask)) return false;
}
int offset;
double value;
int inc = Math.max(r.height / 25, 1);
int mi = 0;
ImageWindow win = imp.getWindow();
if (win != null) win.running = true;
if (measurements == 0) measurements = Analyzer.getMeasurements();
if (showChoice == ELLIPSES) measurements |= ELLIPSE;
measurements &= ~LIMIT; // ignore "Limit to Threshold"
roiNeedsImage =
(measurements & PERIMETER) != 0
|| (measurements & SHAPE_DESCRIPTORS) != 0
|| (measurements & FERET) != 0;
particleCount = 0;
wand = new Wand(ip);
pf = new PolygonFiller();
if (floodFill) {
ImageProcessor ipf = ip.duplicate();
ipf.setValue(fillColor);
ff = new FloodFiller(ipf);
}
roiType = Wand.allPoints() ? Roi.FREEROI : Roi.TRACED_ROI;
for (int y = r.y; y < (r.y + r.height); y++) {
offset = y * width;
for (int x = r.x; x < (r.x + r.width); x++) {
if (pixels != null) value = pixels[offset + x] & 255;
else if (imageType == SHORT) value = ip.getPixel(x, y);
else value = ip.getPixelValue(x, y);
if (value >= level1 && value <= level2) analyzeParticle(x, y, imp, ip);
}
if (showProgress && ((y % inc) == 0)) IJ.showProgress((double) (y - r.y) / r.height);
if (win != null) canceled = !win.running;
if (canceled) {
Macro.abort();
break;
}
}
if (showProgress) IJ.showProgress(1.0);
if (showResults) rt.updateResults();
imp.killRoi();
ip.resetRoi();
ip.reset();
if (displaySummary && IJ.getInstance() != null) updateSliceSummary();
if (addToManager && roiManager != null) roiManager.setEditMode(imp, true);
maxParticleCount = (particleCount > maxParticleCount) ? particleCount : maxParticleCount;
totalCount += particleCount;
if (!canceled) showResults();
return true;
}