public boolean beadCalibration3d() {
imp = IJ.getImage();
if (imp == null) {
IJ.noImage();
return false;
} else if (imp.getStackSize() == 1) {
IJ.error("Stack required");
return false;
} else if (imp.getType() != ImagePlus.GRAY8 && imp.getType() != ImagePlus.GRAY16) {
// In order to support 32bit images, pict[] must be changed to float[], and getPixel(x, y);
// requires a Float.intBitsToFloat() conversion
IJ.error("8 or 16 bit greyscale image required");
return false;
}
width = imp.getWidth();
height = imp.getHeight();
nslices = imp.getStackSize();
imtitle = imp.getTitle();
models[0] = "*None*";
models[1] = "line";
models[2] = "2nd degree polynomial";
models[3] = "3rd degree polynomial";
models[4] = "4th degree polynomial";
GenericDialog gd = new GenericDialog("3D PALM calibration");
gd.addNumericField("Maximum FWHM (in px)", prefs.get("QuickPALM.3Dcal_fwhm", 20), 0);
gd.addNumericField(
"Particle local threshold (% maximum intensity)", prefs.get("QuickPALM.pthrsh", 20), 0);
gd.addNumericField("Z-spacing (nm)", prefs.get("QuickPALM.z-step", 10), 2);
gd.addNumericField("Calibration Z-smoothing (radius)", prefs.get("QuickPALM.window", 1), 0);
gd.addChoice("Model", models, prefs.get("QuickPALM.model", models[3]));
gd.addCheckbox(
"Show divergence of bead positions against model",
prefs.get("QuickPALM.3Dcal_showDivergence", false));
gd.addCheckbox("Show extra particle info", prefs.get("QuickPALM.3Dcal_showExtraInfo", false));
gd.addMessage("\n\nDon't forget to save the table in the end...");
gd.showDialog();
if (gd.wasCanceled()) return false;
fwhm = gd.getNextNumber();
prefs.set("QuickPALM.QuickPALM.3Dcal_fwhm", fwhm);
pthrsh = gd.getNextNumber() / 100;
prefs.set("QuickPALM.pthrsh", pthrsh * 100);
cal_z = gd.getNextNumber();
prefs.set("QuickPALM.z-step", cal_z);
window = (int) gd.getNextNumber();
prefs.set("QuickPALM.window", window);
model = gd.getNextChoice();
prefs.set("QuickPALM.model", model);
part_divergence = gd.getNextBoolean();
prefs.set("QuickPALM.3Dcal_showDivergence", part_divergence);
part_extrainfo = gd.getNextBoolean();
prefs.set("QuickPALM.3Dcal_showExtraInfo", part_extrainfo);
return true;
}
public void run(ImageProcessor ip) {
if (canceled) return;
slice++;
if (imp.getStackSize() > 1 && processStack) imp.setSlice(slice);
if (imp.getType() == ImagePlus.COLOR_RGB) {
ip = (ImageProcessor) imp.getProperty("Mask");
ip.setThreshold(255, 255, ImageProcessor.NO_LUT_UPDATE);
}
if (!analyze(imp, ip)) canceled = true;
if (slice == imp.getStackSize()) {
imp.updateAndDraw();
if (saveRoi != null) imp.setRoi(saveRoi);
}
}
public void doHyperStackProjection(boolean allTimeFrames) {
int start = startSlice;
int stop = stopSlice;
int firstFrame = 1;
int lastFrame = imp.getNFrames();
if (!allTimeFrames) firstFrame = lastFrame = imp.getFrame();
ImageStack stack = new ImageStack(imp.getWidth(), imp.getHeight());
int channels = imp.getNChannels();
int slices = imp.getNSlices();
if (slices == 1) {
slices = imp.getNFrames();
firstFrame = lastFrame = 1;
}
int frames = lastFrame - firstFrame + 1;
increment = channels;
boolean rgb = imp.getBitDepth() == 24;
for (int frame = firstFrame; frame <= lastFrame; frame++) {
for (int channel = 1; channel <= channels; channel++) {
startSlice = (frame - 1) * channels * slices + (start - 1) * channels + channel;
stopSlice = (frame - 1) * channels * slices + (stop - 1) * channels + channel;
if (rgb) doHSRGBProjection(imp);
else doProjection();
stack.addSlice(null, projImage.getProcessor());
}
}
projImage = new ImagePlus(makeTitle(), stack);
projImage.setDimensions(channels, 1, frames);
if (channels > 1) {
projImage = new CompositeImage(projImage, 0);
((CompositeImage) projImage).copyLuts(imp);
if (method == SUM_METHOD || method == SD_METHOD)
((CompositeImage) projImage).resetDisplayRanges();
}
if (frames > 1) projImage.setOpenAsHyperStack(true);
Overlay overlay = imp.getOverlay();
if (overlay != null) {
startSlice = start;
stopSlice = stop;
if (imp.getType() == ImagePlus.COLOR_RGB)
projImage.setOverlay(projectRGBHyperStackRois(overlay));
else projImage.setOverlay(projectHyperStackRois(overlay));
}
IJ.showProgress(1, 1);
}
public void run(String arg) {
Toolkit toolkit = Toolkit.getDefaultToolkit();
Image Image1 = toolkit.getImage(arg);
imp = new ImagePlus("title", Image1);
AffineTransform aTran = new AffineTransform();
if (imp.isComposite()) {
int z = imp.getSlice();
int t = imp.getFrame();
ImagePlus[] channels = split(imp);
imp.changes = false;
imp.setIgnoreFlush(true);
imp.close();
for (int i = 0; i < channels.length; i++) {
channels[i].show();
if (z > 1 || t > 1) channels[i].setPosition(1, z, t);
}
} else if (imp.getType() == ImagePlus.COLOR_RGB) splitRGB(imp);
else IJ.error("Split Channels", "Multichannel image required");
}
/** Splits the specified image into separate channels. */
public static ImagePlus[] split(ImagePlus imp) {
if (imp.getType() == ImagePlus.COLOR_RGB) {
ImageStack[] stacks = splitRGB(imp.getStack(), true);
ImagePlus[] images = new ImagePlus[3];
images[0] = new ImagePlus("red", stacks[0]);
images[1] = new ImagePlus("green", stacks[1]);
images[2] = new ImagePlus("blue", stacks[2]);
return images;
}
int width = imp.getWidth();
int height = imp.getHeight();
int channels = imp.getNChannels();
int slices = imp.getNSlices();
int frames = imp.getNFrames();
int bitDepth = imp.getBitDepth();
int size = slices * frames;
Vector images = new Vector();
HyperStackReducer reducer = new HyperStackReducer(imp);
for (int c = 1; c <= channels; c++) {
ImageStack stack2 = new ImageStack(width, height, size); // create empty stack
stack2.setPixels(
imp.getProcessor().getPixels(), 1); // can't create ImagePlus will null 1st image
ImagePlus imp2 = new ImagePlus("C" + c + "-" + imp.getTitle(), stack2);
stack2.setPixels(null, 1);
imp.setPosition(c, 1, 1);
imp2.setDimensions(1, slices, frames);
imp2.setCalibration(imp.getCalibration());
reducer.reduce(imp2);
if (imp.isComposite() && ((CompositeImage) imp).getMode() == IJ.GRAYSCALE)
IJ.run(imp2, "Grays", "");
if (imp2.getNDimensions() > 3) imp2.setOpenAsHyperStack(true);
images.add(imp2);
}
ImagePlus[] array = new ImagePlus[images.size()];
return (ImagePlus[]) images.toArray(array);
}
void setCalibration(ImagePlus imp) {
if (fi.fileType == FileInfo.GRAY16_SIGNED) {
if (IJ.debugMode) IJ.log("16-bit signed");
double[] coeff = new double[2];
coeff[0] = -32768.0;
coeff[1] = 1.0;
imp.getLocalCalibration().setFunction(Calibration.STRAIGHT_LINE, coeff, "gray value");
}
Properties props = decodeDescriptionString(fi);
Calibration cal = imp.getCalibration();
boolean calibrated = false;
if (fi.pixelWidth > 0.0 && fi.unit != null) {
cal.pixelWidth = fi.pixelWidth;
cal.pixelHeight = fi.pixelHeight;
cal.pixelDepth = fi.pixelDepth;
cal.setUnit(fi.unit);
calibrated = true;
}
if (fi.valueUnit != null) {
int f = fi.calibrationFunction;
if ((f >= Calibration.STRAIGHT_LINE && f <= Calibration.RODBARD2 && fi.coefficients != null)
|| f == Calibration.UNCALIBRATED_OD) {
boolean zeroClip = props != null && props.getProperty("zeroclip", "false").equals("true");
cal.setFunction(f, fi.coefficients, fi.valueUnit, zeroClip);
calibrated = true;
}
}
if (calibrated) checkForCalibrationConflict(imp, cal);
if (fi.frameInterval != 0.0) cal.frameInterval = fi.frameInterval;
if (props == null) return;
cal.xOrigin = getDouble(props, "xorigin");
cal.yOrigin = getDouble(props, "yorigin");
cal.zOrigin = getDouble(props, "zorigin");
cal.info = props.getProperty("info");
cal.fps = getDouble(props, "fps");
cal.loop = getBoolean(props, "loop");
cal.frameInterval = getDouble(props, "finterval");
cal.setTimeUnit(props.getProperty("tunit", "sec"));
double displayMin = getDouble(props, "min");
double displayMax = getDouble(props, "max");
if (!(displayMin == 0.0 && displayMax == 0.0)) {
int type = imp.getType();
ImageProcessor ip = imp.getProcessor();
if (type == ImagePlus.GRAY8 || type == ImagePlus.COLOR_256)
ip.setMinAndMax(displayMin, displayMax);
else if (type == ImagePlus.GRAY16 || type == ImagePlus.GRAY32) {
if (ip.getMin() != displayMin || ip.getMax() != displayMax)
ip.setMinAndMax(displayMin, displayMax);
}
}
int stackSize = imp.getStackSize();
if (stackSize > 1) {
int channels = (int) getDouble(props, "channels");
int slices = (int) getDouble(props, "slices");
int frames = (int) getDouble(props, "frames");
if (channels == 0) channels = 1;
if (slices == 0) slices = 1;
if (frames == 0) frames = 1;
// IJ.log("setCalibration: "+channels+" "+slices+" "+frames);
if (channels * slices * frames == stackSize) {
imp.setDimensions(channels, slices, frames);
if (getBoolean(props, "hyperstack")) imp.setOpenAsHyperStack(true);
}
}
}
/** Restores original disk or network version of image. */
public void revertToSaved(ImagePlus imp) {
Image img;
ProgressBar pb = IJ.getInstance().getProgressBar();
ImageProcessor ip;
String path = fi.directory + fi.fileName;
if (fi.fileFormat == fi.GIF_OR_JPG) {
// restore gif or jpg
img = Toolkit.getDefaultToolkit().createImage(path);
imp.setImage(img);
if (imp.getType() == ImagePlus.COLOR_RGB) Opener.convertGrayJpegTo8Bits(imp);
return;
}
if (fi.fileFormat == fi.DICOM) {
// restore DICOM
ImagePlus imp2 = (ImagePlus) IJ.runPlugIn("ij.plugin.DICOM", path);
if (imp2 != null) imp.setProcessor(null, imp2.getProcessor());
return;
}
if (fi.fileFormat == fi.BMP) {
// restore BMP
ImagePlus imp2 = (ImagePlus) IJ.runPlugIn("ij.plugin.BMP_Reader", path);
if (imp2 != null) imp.setProcessor(null, imp2.getProcessor());
return;
}
if (fi.fileFormat == fi.PGM) {
// restore PGM
ImagePlus imp2 = (ImagePlus) IJ.runPlugIn("ij.plugin.PGM_Reader", path);
if (imp2 != null) imp.setProcessor(null, imp2.getProcessor());
return;
}
if (fi.fileFormat == fi.ZIP_ARCHIVE) {
// restore ".zip" file
ImagePlus imp2 = (new Opener()).openZip(path);
if (imp2 != null) imp.setProcessor(null, imp2.getProcessor());
return;
}
// restore PNG or another image opened using ImageIO
if (fi.fileFormat == fi.IMAGEIO) {
ImagePlus imp2 = (new Opener()).openUsingImageIO(path);
if (imp2 != null) imp.setProcessor(null, imp2.getProcessor());
return;
}
if (fi.nImages > 1) return;
ColorModel cm;
if (fi.url == null || fi.url.equals("")) IJ.showStatus("Loading: " + path);
else IJ.showStatus("Loading: " + fi.url + fi.fileName);
Object pixels = readPixels(fi);
if (pixels == null) return;
cm = createColorModel(fi);
switch (fi.fileType) {
case FileInfo.GRAY8:
case FileInfo.COLOR8:
case FileInfo.BITMAP:
ip = new ByteProcessor(width, height, (byte[]) pixels, cm);
imp.setProcessor(null, ip);
break;
case FileInfo.GRAY16_SIGNED:
case FileInfo.GRAY16_UNSIGNED:
case FileInfo.GRAY12_UNSIGNED:
ip = new ShortProcessor(width, height, (short[]) pixels, cm);
imp.setProcessor(null, ip);
break;
case FileInfo.GRAY32_INT:
case FileInfo.GRAY32_FLOAT:
ip = new FloatProcessor(width, height, (float[]) pixels, cm);
imp.setProcessor(null, ip);
break;
case FileInfo.RGB:
case FileInfo.BGR:
case FileInfo.ARGB:
case FileInfo.ABGR:
case FileInfo.RGB_PLANAR:
img =
Toolkit.getDefaultToolkit()
.createImage(new MemoryImageSource(width, height, (int[]) pixels, 0, width));
imp.setImage(img);
break;
}
}
String getInfo(ImagePlus imp, ImageProcessor ip) {
String s = new String("\n");
s += "Title: " + imp.getTitle() + "\n";
Calibration cal = imp.getCalibration();
int stackSize = imp.getStackSize();
int channels = imp.getNChannels();
int slices = imp.getNSlices();
int frames = imp.getNFrames();
int digits = imp.getBitDepth() == 32 ? 4 : 0;
if (cal.scaled()) {
String unit = cal.getUnit();
String units = cal.getUnits();
s +=
"Width: "
+ IJ.d2s(imp.getWidth() * cal.pixelWidth, 2)
+ " "
+ units
+ " ("
+ imp.getWidth()
+ ")\n";
s +=
"Height: "
+ IJ.d2s(imp.getHeight() * cal.pixelHeight, 2)
+ " "
+ units
+ " ("
+ imp.getHeight()
+ ")\n";
if (slices > 1)
s += "Depth: " + IJ.d2s(slices * cal.pixelDepth, 2) + " " + units + " (" + slices + ")\n";
double xResolution = 1.0 / cal.pixelWidth;
double yResolution = 1.0 / cal.pixelHeight;
int places = Tools.getDecimalPlaces(xResolution, yResolution);
if (xResolution == yResolution)
s += "Resolution: " + IJ.d2s(xResolution, places) + " pixels per " + unit + "\n";
else {
s += "X Resolution: " + IJ.d2s(xResolution, places) + " pixels per " + unit + "\n";
s += "Y Resolution: " + IJ.d2s(yResolution, places) + " pixels per " + unit + "\n";
}
} else {
s += "Width: " + imp.getWidth() + " pixels\n";
s += "Height: " + imp.getHeight() + " pixels\n";
if (stackSize > 1) s += "Depth: " + slices + " pixels\n";
}
if (stackSize > 1)
s +=
"Voxel size: "
+ d2s(cal.pixelWidth)
+ "x"
+ d2s(cal.pixelHeight)
+ "x"
+ d2s(cal.pixelDepth)
+ " "
+ cal.getUnit()
+ "\n";
else
s +=
"Pixel size: "
+ d2s(cal.pixelWidth)
+ "x"
+ d2s(cal.pixelHeight)
+ " "
+ cal.getUnit()
+ "\n";
s += "ID: " + imp.getID() + "\n";
String zOrigin = stackSize > 1 || cal.zOrigin != 0.0 ? "," + d2s(cal.zOrigin) : "";
s += "Coordinate origin: " + d2s(cal.xOrigin) + "," + d2s(cal.yOrigin) + zOrigin + "\n";
int type = imp.getType();
switch (type) {
case ImagePlus.GRAY8:
s += "Bits per pixel: 8 ";
String lut = "LUT";
if (imp.getProcessor().isColorLut()) lut = "color " + lut;
else lut = "grayscale " + lut;
if (imp.isInvertedLut()) lut = "inverting " + lut;
s += "(" + lut + ")\n";
if (imp.getNChannels() > 1) s += displayRanges(imp);
else s += "Display range: " + (int) ip.getMin() + "-" + (int) ip.getMax() + "\n";
break;
case ImagePlus.GRAY16:
case ImagePlus.GRAY32:
if (type == ImagePlus.GRAY16) {
String sign = cal.isSigned16Bit() ? "signed" : "unsigned";
s += "Bits per pixel: 16 (" + sign + ")\n";
} else s += "Bits per pixel: 32 (float)\n";
if (imp.getNChannels() > 1) s += displayRanges(imp);
else {
s += "Display range: ";
double min = ip.getMin();
double max = ip.getMax();
if (cal.calibrated()) {
min = cal.getCValue((int) min);
max = cal.getCValue((int) max);
}
s += IJ.d2s(min, digits) + " - " + IJ.d2s(max, digits) + "\n";
}
break;
case ImagePlus.COLOR_256:
s += "Bits per pixel: 8 (color LUT)\n";
break;
case ImagePlus.COLOR_RGB:
s += "Bits per pixel: 32 (RGB)\n";
break;
}
double interval = cal.frameInterval;
double fps = cal.fps;
if (stackSize > 1) {
ImageStack stack = imp.getStack();
int slice = imp.getCurrentSlice();
String number = slice + "/" + stackSize;
String label = stack.getShortSliceLabel(slice);
if (label != null && label.length() > 0) label = " (" + label + ")";
else label = "";
if (interval > 0.0 || fps != 0.0) {
s += "Frame: " + number + label + "\n";
if (fps != 0.0) {
String sRate =
Math.abs(fps - Math.round(fps)) < 0.00001 ? IJ.d2s(fps, 0) : IJ.d2s(fps, 5);
s += "Frame rate: " + sRate + " fps\n";
}
if (interval != 0.0)
s +=
"Frame interval: "
+ ((int) interval == interval ? IJ.d2s(interval, 0) : IJ.d2s(interval, 5))
+ " "
+ cal.getTimeUnit()
+ "\n";
} else s += "Image: " + number + label + "\n";
if (imp.isHyperStack()) {
if (channels > 1) s += " Channel: " + imp.getChannel() + "/" + channels + "\n";
if (slices > 1) s += " Slice: " + imp.getSlice() + "/" + slices + "\n";
if (frames > 1) s += " Frame: " + imp.getFrame() + "/" + frames + "\n";
}
if (imp.isComposite()) {
if (!imp.isHyperStack() && channels > 1) s += " Channels: " + channels + "\n";
String mode = ((CompositeImage) imp).getModeAsString();
s += " Composite mode: \"" + mode + "\"\n";
}
}
if (ip.getMinThreshold() == ImageProcessor.NO_THRESHOLD) s += "No Threshold\n";
else {
double lower = ip.getMinThreshold();
double upper = ip.getMaxThreshold();
int dp = digits;
if (cal.calibrated()) {
lower = cal.getCValue((int) lower);
upper = cal.getCValue((int) upper);
dp = cal.isSigned16Bit() ? 0 : 4;
}
s += "Threshold: " + IJ.d2s(lower, dp) + "-" + IJ.d2s(upper, dp) + "\n";
}
ImageCanvas ic = imp.getCanvas();
double mag = ic != null ? ic.getMagnification() : 1.0;
if (mag != 1.0) s += "Magnification: " + IJ.d2s(mag, 2) + "\n";
if (cal.calibrated()) {
s += " \n";
int curveFit = cal.getFunction();
s += "Calibration Function: ";
if (curveFit == Calibration.UNCALIBRATED_OD) s += "Uncalibrated OD\n";
else if (curveFit == Calibration.CUSTOM) s += "Custom lookup table\n";
else s += CurveFitter.fList[curveFit] + "\n";
double[] c = cal.getCoefficients();
if (c != null) {
s += " a: " + IJ.d2s(c[0], 6) + "\n";
s += " b: " + IJ.d2s(c[1], 6) + "\n";
if (c.length >= 3) s += " c: " + IJ.d2s(c[2], 6) + "\n";
if (c.length >= 4) s += " c: " + IJ.d2s(c[3], 6) + "\n";
if (c.length >= 5) s += " c: " + IJ.d2s(c[4], 6) + "\n";
}
s += " Unit: \"" + cal.getValueUnit() + "\"\n";
} else s += "Uncalibrated\n";
FileInfo fi = imp.getOriginalFileInfo();
if (fi != null) {
if (fi.url != null && !fi.url.equals("")) s += "URL: " + fi.url + "\n";
else if (fi.directory != null && fi.fileName != null)
s += "Path: " + fi.directory + fi.fileName + "\n";
}
ImageWindow win = imp.getWindow();
if (win != null) {
Point loc = win.getLocation();
Dimension screen = IJ.getScreenSize();
s +=
"Screen location: "
+ loc.x
+ ","
+ loc.y
+ " ("
+ screen.width
+ "x"
+ screen.height
+ ")\n";
}
Overlay overlay = imp.getOverlay();
if (overlay != null) {
String hidden = imp.getHideOverlay() ? " (hidden)" : " ";
int n = overlay.size();
String elements = n == 1 ? " element" : " elements";
s += "Overlay: " + n + elements + (imp.getHideOverlay() ? " (hidden)" : "") + "\n";
} else s += "No Overlay\n";
Roi roi = imp.getRoi();
if (roi == null) {
if (cal.calibrated()) s += " \n";
s += "No Selection\n";
} else if (roi instanceof EllipseRoi) {
s += "\nElliptical Selection\n";
double[] p = ((EllipseRoi) roi).getParams();
double dx = p[2] - p[0];
double dy = p[3] - p[1];
double major = Math.sqrt(dx * dx + dy * dy);
s += " Major: " + IJ.d2s(major, 2) + "\n";
s += " Minor: " + IJ.d2s(major * p[4], 2) + "\n";
s += " X1: " + IJ.d2s(p[0], 2) + "\n";
s += " Y1: " + IJ.d2s(p[1], 2) + "\n";
s += " X2: " + IJ.d2s(p[2], 2) + "\n";
s += " Y2: " + IJ.d2s(p[3], 2) + "\n";
s += " Aspect ratio: " + IJ.d2s(p[4], 2) + "\n";
} else {
s += " \n";
s += roi.getTypeAsString() + " Selection";
String points = null;
if (roi instanceof PointRoi) {
int npoints = ((PolygonRoi) roi).getNCoordinates();
String suffix = npoints > 1 ? "s)" : ")";
points = " (" + npoints + " point" + suffix;
}
String name = roi.getName();
if (name != null) {
s += " (\"" + name + "\")";
if (points != null) s += "\n " + points;
} else if (points != null) s += points;
s += "\n";
Rectangle r = roi.getBounds();
if (roi instanceof Line) {
Line line = (Line) roi;
s += " X1: " + IJ.d2s(line.x1d * cal.pixelWidth) + "\n";
s += " Y1: " + IJ.d2s(yy(line.y1d, imp) * cal.pixelHeight) + "\n";
s += " X2: " + IJ.d2s(line.x2d * cal.pixelWidth) + "\n";
s += " Y2: " + IJ.d2s(yy(line.y2d, imp) * cal.pixelHeight) + "\n";
} else if (cal.scaled()) {
s += " X: " + IJ.d2s(cal.getX(r.x)) + " (" + r.x + ")\n";
s += " Y: " + IJ.d2s(cal.getY(r.y, imp.getHeight())) + " (" + r.y + ")\n";
s += " Width: " + IJ.d2s(r.width * cal.pixelWidth) + " (" + r.width + ")\n";
s += " Height: " + IJ.d2s(r.height * cal.pixelHeight) + " (" + r.height + ")\n";
} else {
s += " X: " + r.x + "\n";
s += " Y: " + yy(r.y, imp) + "\n";
s += " Width: " + r.width + "\n";
s += " Height: " + r.height + "\n";
}
}
return s;
}
/**
* 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;
}
ImageProcessor setup(ImagePlus imp) {
ImageProcessor ip;
int type = imp.getType();
if (type != ImagePlus.COLOR_RGB) return null;
ip = imp.getProcessor();
int id = imp.getID();
int slice = imp.getCurrentSlice();
if ((id != previousImageID) | (slice != previousSlice) | (flag)) {
flag = false; // if true, flags a change from HSB to RGB or viceversa
numSlices = imp.getStackSize();
stack = imp.getStack();
width = stack.getWidth();
height = stack.getHeight();
numPixels = width * height;
hSource = new byte[numPixels];
sSource = new byte[numPixels];
bSource = new byte[numPixels];
// restore = (int[])ip.getPixelsCopy(); //This runs into trouble sometimes, so do it the
// long way:
int[] temp = (int[]) ip.getPixels();
restore = new int[numPixels];
for (int i = 0; i < numPixels; i++) restore[i] = temp[i];
fillMask = new int[numPixels];
// Get hsb or rgb from image.
ColorProcessor cp = (ColorProcessor) ip;
IJ.showStatus("Gathering data");
if (isRGB) cp.getRGB(hSource, sSource, bSource);
else cp.getHSB(hSource, sSource, bSource);
IJ.showStatus("done");
// Create a spectrum ColorModel for the Hue histogram plot.
Color c;
byte[] reds = new byte[256];
byte[] greens = new byte[256];
byte[] blues = new byte[256];
for (int i = 0; i < 256; i++) {
c = Color.getHSBColor(i / 255f, 1f, 1f);
reds[i] = (byte) c.getRed();
greens[i] = (byte) c.getGreen();
blues[i] = (byte) c.getBlue();
}
ColorModel cm = new IndexColorModel(8, 256, reds, greens, blues);
// Make an image with just the hue from the RGB image and the spectrum LUT.
// This is just for a hue histogram for the plot. Do not show it.
// ByteProcessor bpHue = new ByteProcessor(width,height,h,cm);
ByteProcessor bpHue = new ByteProcessor(width, height, hSource, cm);
ImagePlus impHue = new ImagePlus("Hue", bpHue);
// impHue.show();
ByteProcessor bpSat = new ByteProcessor(width, height, sSource, cm);
ImagePlus impSat = new ImagePlus("Sat", bpSat);
// impSat.show();
ByteProcessor bpBri = new ByteProcessor(width, height, bSource, cm);
ImagePlus impBri = new ImagePlus("Bri", bpBri);
// impBri.show();
plot.setHistogram(impHue, 0);
splot.setHistogram(impSat, 1);
bplot.setHistogram(impBri, 2);
updateLabels();
updatePlot();
updateScrollBars();
imp.updateAndDraw();
}
previousImageID = id;
previousSlice = slice;
return ip;
}
public void Calc_5Fr(ImagePlus imp1, ImagePlus imp2) {
if (imp1.getType() != imp2.getType()) {
error();
return;
}
if (imp1.getType() == 0) { // getType returns 0 for 8-bit, 1 for 16-bit
bitDepth = "8-bit";
Prefs.set("ps.bitDepth", bitDepth);
} else {
bitDepth = "16-bit";
Prefs.set("ps.bitDepth", bitDepth);
}
int width = imp1.getWidth();
int height = imp1.getHeight();
if (width != imp2.getWidth() || height != imp2.getHeight()) {
error();
return;
}
ImageStack stack1 = imp1.getStack();
// if (bgStackTitle != "NoBg") ImageStack stack2 = imp2.getStack();
ImageStack stack2 = imp2.getStack();
ImageProcessor ip = imp1.getProcessor();
int dimension = width * height;
byte[] pixB;
short[] pixS;
float[][] pixF = new float[5][dimension];
float[][] pixFBg = new float[5][dimension];
float a;
float b;
float den;
float aSmp;
float bSmp;
float denSmp;
float aBg;
float bBg;
float denBg;
float retF;
float azimF;
byte[] retB = new byte[dimension];
short[] retS = new short[dimension];
byte[] azimB = new byte[dimension];
short[] azimS = new short[dimension];
// Derived Variables:
float swingAngle = 2f * (float) Math.PI * swing;
float tanSwingAngleDiv2 = (float) Math.tan(swingAngle / 2.f);
float tanSwingAngleDiv2DivSqrt2 = (float) (Math.tan(swingAngle / 2.f) / Math.sqrt(2));
float wavelengthDiv2Pi = wavelength / (2f * (float) Math.PI);
// get the pixels of each slice in the stack and convert to float
for (int i = 0; i < 5; i++) {
if (bitDepth == "8-bit") {
pixB = (byte[]) stack1.getPixels(i + 3);
for (int j = 0; j < dimension; j++) pixF[i][j] = 0xff & pixB[j];
if (bgStackTitle != "NoBg") {
pixB = (byte[]) stack2.getPixels(i + 3);
for (int j = 0; j < dimension; j++) pixFBg[i][j] = 0xff & pixB[j];
}
} else {
pixS = (short[]) stack1.getPixels(i + 3);
for (int j = 0; j < dimension; j++) pixF[i][j] = (float) pixS[j];
if (bgStackTitle != "NoBg") {
pixS = (short[]) stack2.getPixels(i + 3);
for (int j = 0; j < dimension; j++) pixFBg[i][j] = (float) pixS[j];
}
}
}
// Algorithm
// terms a and b
for (int j = 0; j < dimension; j++) {
denSmp = (pixF[1][j] + pixF[2][j] + pixF[3][j] + pixF[4][j] - 4 * pixF[0][j]) / 2;
denBg = denSmp;
a = (pixF[4][j] - pixF[1][j]);
aSmp = a;
aBg = a;
b = (pixF[2][j] - pixF[3][j]);
bSmp = b;
bBg = b;
if (bgStackTitle != "NoBg") {
denBg = (pixFBg[1][j] + pixFBg[2][j] + pixFBg[3][j] + pixFBg[4][j] - 4 * pixFBg[0][j]) / 2;
aBg = pixFBg[4][j] - pixFBg[1][j];
bBg = pixFBg[2][j] - pixFBg[3][j];
}
// Special case of sample retardance half wave, denSmp = 0
if (denSmp == 0) {
retF = (float) wavelength / 4;
azimF =
(float) (a == 0 & b == 0 ? 0 : (azimRef + 90 + 90 * Math.atan2(a, b) / Math.PI) % 180);
} else {
// Retardance, the background correction can be improved by separately considering sample
// retardance values larger than a quarter wave
if (bgStackTitle != "NoBg") {
a = aSmp / denSmp - aBg / denBg;
b = bSmp / denSmp - bBg / denBg;
} else {
a = aSmp / denSmp;
b = bSmp / denSmp;
}
retF = (float) Math.atan(tanSwingAngleDiv2 * Math.sqrt(a * a + b * b));
if (denSmp < 0) retF = (float) Math.PI - retF;
retF = retF * wavelengthDiv2Pi; // convert to nm
if (retF > retCeiling) retF = retCeiling;
// Orientation
if ((bgStackTitle == "NoBg") || ((bgStackTitle != "NoBg") && (Math.abs(denSmp) < 1))) {
a = aSmp;
b = bSmp;
}
azimF =
(float) (a == 0 & b == 0 ? 0 : (azimRef + 90 + 90 * Math.atan2(a, b) / Math.PI) % 180);
}
if (bitDepth == "8-bit") retB[j] = (byte) (((int) (255 * retF / retCeiling)) & 0xff);
else retS[j] = (short) (4095 * retF / retCeiling);
if (mirror == "Yes") azimF = 180 - azimF;
if (bitDepth == "8-bit") azimB[j] = (byte) (((int) azimF) & 0xff);
else azimS[j] = (short) (azimF * 10f);
}
// show the resulting images in slice 1 and 2
imp1.setSlice(3);
if (bitDepth == "8-bit") {
stack1.setPixels(retB, 1);
stack1.setPixels(azimB, 2);
} else {
stack1.setPixels(retS, 1);
stack1.setPixels(azimS, 2);
}
imp1.setSlice(1);
IJ.selectWindow(imp1.getTitle());
Prefs.set("ps.sampleStackTitle", sampleStackTitle);
Prefs.set("ps.bgStackTitle", bgStackTitle);
Prefs.set("ps.mirror", mirror);
Prefs.set("ps.wavelength", wavelength);
Prefs.set("ps.swing", swing);
Prefs.set("ps.retCeiling", retCeiling);
Prefs.set("ps.azimRef", azimRef);
Prefs.savePreferences();
}
/*------------------------------------------------------------------*/
public void run(String arg) {
ImagePlus imp = WindowManager.getCurrentImage();
this.imp = imp;
if (imp == null) {
IJ.noImage();
return;
}
if ((1 < imp.getStackSize()) && (imp.getType() == imp.COLOR_256)) {
IJ.error("Stack of color images not supported (use grayscale)");
return;
}
if (1 < imp.getStackSize()) {
if (imp.getStack().isRGB()) {
IJ.error("RGB color images not supported (use grayscale)");
return;
}
}
if (1 < imp.getStackSize()) {
if (imp.getStack().isHSB()) {
IJ.error("HSB color images not supported (use grayscale)");
return;
}
}
if (imp.getType() == imp.COLOR_256) {
IJ.error("Indexed color images not supported (use grayscale)");
return;
}
if (imp.getType() == imp.COLOR_RGB) {
IJ.error("Color images not supported (use grayscale)");
return;
}
differentialsDialog dialog =
new differentialsDialog(IJ.getInstance(), "Differentials", true, operation);
GUI.center(dialog);
dialog.setVisible(true);
cancel = dialog.getCancel();
operation = dialog.getOperation();
dialog.dispose();
if (cancel) {
return;
}
imp.startTiming();
if (1 < imp.getStackSize()) {
if (!(imp.getProcessor().getPixels() instanceof float[])) {
new StackConverter(imp).convertToGray32();
}
} else {
if (!(imp.getProcessor().getPixels() instanceof float[])) {
new ImageConverter(imp).convertToGray32();
}
}
ImageStack stack = imp.getStack();
stackSize = stack.getSize();
Undo.reset();
setupProgressBar();
resetProgressBar();
for (int i = 1; (i <= stackSize); i++) {
ImageProcessor ip = stack.getProcessor(i);
doIt(ip);
imp.getProcessor().resetMinAndMax();
imp.setSlice(i);
imp.updateAndRepaintWindow();
}
imp.getProcessor().resetMinAndMax();
imp.setSlice(1);
imp.updateAndRepaintWindow();
cleanUpProgressBar();
IJ.showTime(imp, imp.getStartTime(), "Differentials: ");
ImageWindow win = imp.getWindow();
if (win != null) {
win.running = false;
}
} /* end run */
public void run(String arg) {
imp = IJ.getImage();
int stackSize = imp.getStackSize();
if (imp == null) {
IJ.noImage();
return;
}
// Make sure input image is a stack.
if (stackSize == 1) {
IJ.error("Z Project", "Stack required");
return;
}
// Check for inverting LUT.
if (imp.getProcessor().isInvertedLut()) {
if (!IJ.showMessageWithCancel("ZProjection", lutMessage)) return;
}
// Set default bounds.
int channels = imp.getNChannels();
int frames = imp.getNFrames();
int slices = imp.getNSlices();
isHyperstack =
imp.isHyperStack()
|| (ij.macro.Interpreter.isBatchMode()
&& ((frames > 1 && frames < stackSize) || (slices > 1 && slices < stackSize)));
boolean simpleComposite = channels == stackSize;
if (simpleComposite) isHyperstack = false;
startSlice = 1;
if (isHyperstack) {
int nSlices = imp.getNSlices();
if (nSlices > 1) stopSlice = nSlices;
else stopSlice = imp.getNFrames();
} else stopSlice = stackSize;
// Build control dialog
GenericDialog gd = buildControlDialog(startSlice, stopSlice);
gd.showDialog();
if (gd.wasCanceled()) return;
if (!imp.lock()) return; // exit if in use
long tstart = System.currentTimeMillis();
setStartSlice((int) gd.getNextNumber());
setStopSlice((int) gd.getNextNumber());
method = gd.getNextChoiceIndex();
Prefs.set(METHOD_KEY, method);
if (isHyperstack) {
allTimeFrames = imp.getNFrames() > 1 && imp.getNSlices() > 1 ? gd.getNextBoolean() : false;
doHyperStackProjection(allTimeFrames);
} else if (imp.getType() == ImagePlus.COLOR_RGB) doRGBProjection(true);
else doProjection(true);
if (arg.equals("") && projImage != null) {
long tstop = System.currentTimeMillis();
projImage.setCalibration(imp.getCalibration());
if (simpleComposite) IJ.run(projImage, "Grays", "");
projImage.show("ZProjector: " + IJ.d2s((tstop - tstart) / 1000.0, 2) + " seconds");
}
imp.unlock();
IJ.register(ZProjector.class);
return;
}
