예제 #1
0
  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;
  }
예제 #2
0
    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;
    }
예제 #3
0
  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();
  }