/* Method takes roi and substract its mean value from each frame in the image*/ public static ImagePlus DarkNoiseRemoval(ImagePlus imp_in, Roi noise_roi) { ImagePlus imp_out = imp_in.duplicate(); int size = imp_in.getImageStackSize(); ImageProcessor ip = imp_out.getProcessor(); double minThreshold = ip.getMinThreshold(); double maxThreshold = ip.getMaxThreshold(); Calibration cal = imp_in.getCalibration(); int measurements = Analyzer.getMeasurements(); int current = imp_in.getCurrentSlice(); ImageStack stack = imp_in.getStack(); for (int i = 1; i <= size; i++) { ip = stack.getProcessor(i); if (minThreshold != ImageProcessor.NO_THRESHOLD) ip.setThreshold(minThreshold, maxThreshold, ImageProcessor.NO_LUT_UPDATE); ip.setRoi(noise_roi); ImageStatistics stats = ImageStatistics.getStatistics(ip, measurements, cal); ip.resetRoi(); ip.subtract(stats.mean); stack.setProcessor(ip, i); } imp_out.setStack(stack); return imp_out; }
/** * 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; }
/** Displays a modal options dialog. */ public boolean showDialog() { Calibration cal = imp != null ? imp.getCalibration() : (new Calibration()); double unitSquared = cal.pixelWidth * cal.pixelHeight; if (pixelUnits) unitSquared = 1.0; if (Macro.getOptions() != null) { boolean oldMacro = updateMacroOptions(); if (oldMacro) unitSquared = 1.0; staticMinSize = 0.0; staticMaxSize = DEFAULT_MAX_SIZE; staticMinCircularity = 0.0; staticMaxCircularity = 1.0; staticShowChoice = NOTHING; } GenericDialog gd = new GenericDialog("Analyze Particles"); minSize = staticMinSize; maxSize = staticMaxSize; minCircularity = staticMinCircularity; maxCircularity = staticMaxCircularity; showChoice = staticShowChoice; if (maxSize == 999999) maxSize = DEFAULT_MAX_SIZE; options = staticOptions; String unit = cal.getUnit(); boolean scaled = cal.scaled(); if (unit.equals("inch")) { unit = "pixel"; unitSquared = 1.0; scaled = false; pixelUnits = true; } String units = unit + "^2"; int places = 0; double cmin = minSize * unitSquared; if ((int) cmin != cmin) places = 2; double cmax = maxSize * unitSquared; if ((int) cmax != cmax && cmax != DEFAULT_MAX_SIZE) places = 2; String minStr = ResultsTable.d2s(cmin, places); if (minStr.indexOf("-") != -1) { for (int i = places; i <= 6; i++) { minStr = ResultsTable.d2s(cmin, i); if (minStr.indexOf("-") == -1) break; } } String maxStr = ResultsTable.d2s(cmax, places); if (maxStr.indexOf("-") != -1) { for (int i = places; i <= 6; i++) { maxStr = ResultsTable.d2s(cmax, i); if (maxStr.indexOf("-") == -1) break; } } if (scaled) gd.setInsets(5, 0, 0); gd.addStringField("Size (" + units + "):", minStr + "-" + maxStr, 12); if (scaled) { gd.setInsets(0, 40, 5); gd.addCheckbox("Pixel units", pixelUnits); } gd.addStringField("Circularity:", IJ.d2s(minCircularity) + "-" + IJ.d2s(maxCircularity), 12); gd.addChoice("Show:", showStrings, showStrings[showChoice]); String[] labels = new String[8]; boolean[] states = new boolean[8]; labels[0] = "Display results"; states[0] = (options & SHOW_RESULTS) != 0; labels[1] = "Exclude on edges"; states[1] = (options & EXCLUDE_EDGE_PARTICLES) != 0; labels[2] = "Clear results"; states[2] = (options & CLEAR_WORKSHEET) != 0; labels[3] = "Include holes"; states[3] = (options & INCLUDE_HOLES) != 0; labels[4] = "Summarize"; states[4] = (options & DISPLAY_SUMMARY) != 0; labels[5] = "Record starts"; states[5] = (options & RECORD_STARTS) != 0; labels[6] = "Add to Manager"; states[6] = (options & ADD_TO_MANAGER) != 0; labels[7] = "In_situ Show"; states[7] = (options & IN_SITU_SHOW) != 0; gd.addCheckboxGroup(4, 2, labels, states); gd.addHelp(IJ.URL + "/docs/menus/analyze.html#ap"); gd.showDialog(); if (gd.wasCanceled()) return false; String size = gd.getNextString(); // min-max size if (scaled) pixelUnits = gd.getNextBoolean(); if (pixelUnits) unitSquared = 1.0; else unitSquared = cal.pixelWidth * cal.pixelHeight; String[] minAndMax = Tools.split(size, " -"); double mins = gd.parseDouble(minAndMax[0]); double maxs = minAndMax.length == 2 ? gd.parseDouble(minAndMax[1]) : Double.NaN; minSize = Double.isNaN(mins) ? DEFAULT_MIN_SIZE : mins / unitSquared; maxSize = Double.isNaN(maxs) ? DEFAULT_MAX_SIZE : maxs / unitSquared; if (minSize < DEFAULT_MIN_SIZE) minSize = DEFAULT_MIN_SIZE; if (maxSize < minSize) maxSize = DEFAULT_MAX_SIZE; staticMinSize = minSize; staticMaxSize = maxSize; minAndMax = Tools.split(gd.getNextString(), " -"); // min-max circularity double minc = gd.parseDouble(minAndMax[0]); double maxc = minAndMax.length == 2 ? gd.parseDouble(minAndMax[1]) : Double.NaN; minCircularity = Double.isNaN(minc) ? 0.0 : minc; maxCircularity = Double.isNaN(maxc) ? 1.0 : maxc; if (minCircularity < 0.0 || minCircularity > 1.0) minCircularity = 0.0; if (maxCircularity < minCircularity || maxCircularity > 1.0) maxCircularity = 1.0; if (minCircularity == 1.0 && maxCircularity == 1.0) minCircularity = 0.0; staticMinCircularity = minCircularity; staticMaxCircularity = maxCircularity; if (gd.invalidNumber()) { IJ.error("Bins invalid."); canceled = true; return false; } showChoice = gd.getNextChoiceIndex(); staticShowChoice = showChoice; if (gd.getNextBoolean()) options |= SHOW_RESULTS; else options &= ~SHOW_RESULTS; if (gd.getNextBoolean()) options |= EXCLUDE_EDGE_PARTICLES; else options &= ~EXCLUDE_EDGE_PARTICLES; if (gd.getNextBoolean()) options |= CLEAR_WORKSHEET; else options &= ~CLEAR_WORKSHEET; if (gd.getNextBoolean()) options |= INCLUDE_HOLES; else options &= ~INCLUDE_HOLES; if (gd.getNextBoolean()) options |= DISPLAY_SUMMARY; else options &= ~DISPLAY_SUMMARY; if (gd.getNextBoolean()) options |= RECORD_STARTS; else options &= ~RECORD_STARTS; if (gd.getNextBoolean()) options |= ADD_TO_MANAGER; else options &= ~ADD_TO_MANAGER; if (gd.getNextBoolean()) options |= IN_SITU_SHOW; else options &= ~IN_SITU_SHOW; staticOptions = options; options |= SHOW_PROGRESS; if ((options & DISPLAY_SUMMARY) != 0) Analyzer.setMeasurements(Analyzer.getMeasurements() | AREA); return true; }