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;
}
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();
}