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;
}
private boolean showDialog() {
String[] types = {"RAW", "JPEG", "ZLIB"};
GenericDialog gd = new GenericDialog("Generate Bricks");
gd.addChoice("FileType", types, filetype);
gd.addNumericField("JPEG quality", jpeg_quality, 0);
gd.addNumericField("Max file size (MB)", bdsizelimit, 0);
int[] wlist = WindowManager.getIDList();
if (wlist == null) return false;
String[] titles = new String[wlist.length];
for (int i = 0; i < wlist.length; i++) titles[i] = "";
int tnum = 0;
for (int i = 0; i < wlist.length; i++) {
ImagePlus imp = WindowManager.getImage(wlist[i]);
if (imp != null) {
titles[tnum] = imp.getTitle();
tnum++;
}
}
gd.addChoice("Source image: ", titles, titles[0]);
gd.showDialog();
if (gd.wasCanceled()) return false;
filetype = types[gd.getNextChoiceIndex()];
jpeg_quality = (int) gd.getNextNumber();
if (jpeg_quality > 100) jpeg_quality = 100;
if (jpeg_quality < 0) jpeg_quality = 0;
bdsizelimit = (int) gd.getNextNumber();
int id = gd.getNextChoiceIndex();
lvImgTitle = new ArrayList<String>();
lvImgTitle.add(titles[id]);
Prefs.set("filetype.string", filetype);
Prefs.set("jpeg_quality.int", jpeg_quality);
Prefs.set("bdsizelimit.int", bdsizelimit);
return true;
}
public void run(String arg) {
int[] wList = WindowManager.getIDList();
if (wList == null) {
IJ.error("No images are open.");
return;
}
double thalf = 0.5;
boolean keep;
GenericDialog gd = new GenericDialog("Bleach correction");
gd.addNumericField("t½:", thalf, 1);
gd.addCheckbox("Keep source stack:", true);
gd.showDialog();
if (gd.wasCanceled()) return;
long start = System.currentTimeMillis();
thalf = gd.getNextNumber();
keep = gd.getNextBoolean();
if (keep) IJ.run("Duplicate...", "title='Bleach corrected' duplicate");
ImagePlus imp1 = WindowManager.getCurrentImage();
int d1 = imp1.getStackSize();
double v1, v2;
int width = imp1.getWidth();
int height = imp1.getHeight();
ImageProcessor ip1, ip2, ip3;
int slices = imp1.getStackSize();
ImageStack stack1 = imp1.getStack();
ImageStack stack2 = imp1.getStack();
int currentSlice = imp1.getCurrentSlice();
for (int n = 1; n <= slices; n++) {
ip1 = stack1.getProcessor(n);
ip3 = stack1.getProcessor(1);
ip2 = stack2.getProcessor(n);
for (int x = 0; x < width; x++) {
for (int y = 0; y < height; y++) {
v1 = ip1.getPixelValue(x, y);
v2 = ip3.getPixelValue(x, y);
// =B8/(EXP(-C$7*A8))
v1 = (v1 / Math.exp(-n * thalf));
ip2.putPixelValue(x, y, v1);
}
}
IJ.showProgress((double) n / slices);
IJ.showStatus(n + "/" + slices);
}
// stack2.show();
imp1.updateAndDraw();
}
public boolean dialogItemChanged(GenericDialog gd, AWTEvent e) {
checkc2 = gd.getNextBoolean();
double[] params = new double[10];
int[] fixes = new int[10];
for (int i = 0; i < params.length; i++) {
params[i] = gd.getNextNumber();
if (gd.getNextBoolean()) {
fixes[i] = 1;
} else {
fixes[i] = 0;
}
}
gd.getNextBoolean();
gd.getNextNumber();
gd.getNextNumber();
NLLSfit_v2 fitclass = new NLLSfit_v2(this, 0);
double[] stats = new double[2];
float[] fit = fitclass.fitdata(params, fixes, null, tempdata, null, stats, true);
pw.updateSeries(fit, series, false);
c2 = (float) stats[1];
return true;
}
public void run(String arg) {
ImagePlus imp = WindowManager.getCurrentImage();
if (imp == null) {
IJ.noImage();
return;
}
if (imp.getStackSize() > 1) {
IJ.error("This command requires a montage");
return;
}
GenericDialog gd = new GenericDialog("Stack Maker");
gd.addNumericField("Images_per_row: ", w, 0);
gd.addNumericField("Images_per_column: ", h, 0);
gd.addNumericField("Border width: ", b, 0);
gd.showDialog();
if (gd.wasCanceled()) return;
w = (int) gd.getNextNumber();
h = (int) gd.getNextNumber();
b = (int) gd.getNextNumber();
ImageStack stack = makeStack(imp.getProcessor(), w, h, b);
new ImagePlus("Stack", stack).show();
}
boolean showoptions(double[] params, int[] fixes) {
// GenericDialog gd=new NonBlockingGenericDialog("Options");
GenericDialog gd = new GenericDialog("Options");
gd.addCheckbox("Check Chi Squared", checkc2);
for (int i = 0; i < 10; i++) {
gd.addNumericField("P" + (i + 1), params[i], 5, 10, null);
gd.addCheckbox("Fix?", (fixes[i] == 1));
}
gd.addCheckbox("Get_Errors", false);
gd.addCheckbox("Set_Constraints", false);
gd.addNumericField("Iterations", iterations, 0, 10, null);
gd.addNumericField("chi squared", c2, 5, 10, null);
gd.addDialogListener(this);
gd.showDialog();
if (gd.wasCanceled()) {
return false;
}
checkc2 = gd.getNextBoolean();
for (int i = 0; i < 10; i++) {
params[i] = gd.getNextNumber();
if (gd.getNextBoolean()) {
fixes[i] = 1;
} else {
fixes[i] = 0;
}
}
boolean geterrors = gd.getNextBoolean();
boolean setconstraints = gd.getNextBoolean();
for (int i = 0; i < 10; i++) {
if (function.indexOf("P" + (i + 1)) < 0) {
fixes[i] = 1;
}
}
if (geterrors) {
if (!get_errors(params, fixes)) {
return false;
}
}
if (setconstraints) constraints = get_constraints(params);
return true;
}
public double[][] get_constraints(double[] params) {
// here we populate the constraints
GenericDialog gd = new GenericDialog("Constraints");
for (int i = 0; i < 10; i++) {
if (constraints == null) {
gd.addNumericField("P" + (i + 1) + "_upper", params[i], 5, 10, null);
gd.addNumericField("P" + (i + 1) + "_lower", params[i], 5, 10, null);
} else {
gd.addNumericField("P" + (i + 1) + "_upper", constraints[1][i], 5, 10, null);
gd.addNumericField("P" + (i + 1) + "_lower", constraints[0][i], 5, 10, null);
}
}
gd.showDialog();
if (gd.wasCanceled()) {
return null;
}
double[][] constraints = new double[2][10];
for (int i = 0; i < 10; i++) {
constraints[1][i] = gd.getNextNumber();
constraints[0][i] = gd.getNextNumber();
}
return constraints;
}
public boolean dialogItemChanged(GenericDialog gd, AWTEvent e) {
int width = imp.getWidth();
int height = imp.getHeight();
type = gd.getNextChoice();
areaPerPoint = gd.getNextNumber();
color = gd.getNextChoice();
randomOffset = gd.getNextBoolean();
double minArea = (width * height) / 50000.0;
if (type.equals(types[1]) && minArea < 144.0) minArea = 144.0;
else if (minArea < 16) minArea = 16.0;
if (areaPerPoint / (pixelWidth * pixelHeight) < minArea) {
String err = "\"Area per Point\" too small";
if (gd.wasOKed()) IJ.error("Grid", err);
else IJ.showStatus(err);
return true;
}
double tileSize = Math.sqrt(areaPerPoint);
tileWidth = tileSize / pixelWidth;
tileHeight = tileSize / pixelHeight;
if (randomOffset) {
xstart = (int) (random.nextDouble() * tileWidth);
ystart = (int) (random.nextDouble() * tileHeight);
} else {
xstart = (int) (tileWidth / 2.0 + 0.5);
ystart = (int) (tileHeight / 2.0 + 0.5);
}
linesV = (int) ((width - xstart) / tileWidth) + 1;
linesH = (int) ((height - ystart) / tileHeight) + 1;
if (gd.invalidNumber()) return true;
if (type.equals(types[0])) drawLines();
else if (type.equals(types[1])) drawCrosses();
else if (type.equals(types[2])) drawPoints();
else showGrid(null);
return true;
}
public void run(String arg) {
int[] wList = WindowManager.getIDList();
if (wList==null) {
IJ.error("No images are open.");
return;
}
double kernel=3;
double kernelsum = 0;
double kernelvarsum =0;
double kernalvar = 0;
double sigmawidth = 2;
int kernelindex, minpixnumber;
String[] kernelsize = { "3�,"5�, "7�, "9�};
GenericDialog gd = new GenericDialog("Sigma Filter");
gd.addChoice("Kernel size", kernelsize, kernelsize[0]);
gd.addNumericField("Sigma width",sigmawidth , 2);
gd.addNumericField("Minimum number of pixels", 1, 0);
gd.addCheckbox("Keep source:",true);
gd.addCheckbox("Do all stack:",true);
gd.addCheckbox("Modified Lee's FIlter:",true);
gd.showDialog();
if (gd.wasCanceled()) return ;
kernelindex = gd.getNextChoiceIndex();
sigmawidth = gd.getNextNumber();
minpixnumber = ((int)gd.getNextNumber());
boolean keep = gd.getNextBoolean();
boolean doallstack = gd.getNextBoolean();
boolean modified = gd.getNextBoolean();
if (kernelindex==0) kernel = 3;
if (kernelindex==1) kernel = 5;
if (kernelindex==2) kernel = 7;
if (kernelindex==3) kernel = 9;
long start = System.currentTimeMillis();
if (minpixnumber> (kernel*kernel)){
IJ.showMessage("Sigma filter", "There must be more pixels in the kernel than+\n" + "the minimum number to be included");
return;
}
double v, midintensity;
int x, y, ix, iy;
double sum = 0;
double backupsum =0;
int count = 0;
int n = 0;
if (keep) {IJ.run("Select All"); IJ.run("Duplicate...", "title='Sigma filtered' duplicate");}
int radius = (int)(kernel-1)/2;
ImagePlus imp = WindowManager.getCurrentImage();
ImageStack stack1 = imp.getStack();
int width = imp.getWidth();
int height = imp.getHeight();
int nslices = stack1.getSize();
int cslice = imp.getCurrentSlice();
double status = width*height*nslices;
ImageProcessor ip = imp.getProcessor();
int sstart = 1;
if (!doallstack) {sstart = cslice; nslices=sstart;status = status/nslices;};
for (int i=sstart; i<=nslices; i++) {
imp.setSlice(i);
for (x=radius;x<width+radius;x++) {
for (y=radius;y<height+radius;y++) {
midintensity = ip.getPixelValue(x,y);
count = 0;
sum = 0;
kernelsum =0;
kernalvar =0;
kernelvarsum =0;
backupsum = 0;
//calculate mean of kernel value
for (ix=0;ix<kernel;ix++) {
for (iy=0;iy<kernel;iy++) {
v = ip.getPixelValue(x+ix-radius,y+iy-radius);
kernelsum = kernelsum+v;
}
}
double sigmacalcmean = (kernelsum/(kernel*kernel));
//calculate variance of kernel
for (ix=0;ix<kernel;ix++) {
for (iy=0;iy<kernel;iy++) {
v = ip.getPixelValue(x+ix-radius,y+iy-radius);
kernalvar = (v-sigmacalcmean)*(v-sigmacalcmean);
kernelvarsum = kernelvarsum + kernalvar;
}
}
//double variance = kernelvarsum/kernel;
double sigmacalcvar = kernelvarsum/((kernel*kernel)-1);
//calcuate sigma range = sqrt(variance/(mean^2)) � sigmawidth
double sigmarange = sigmawidth*(Math.sqrt((sigmacalcvar) /(sigmacalcmean*sigmacalcmean)));
//calulate sigma top value and bottom value
double sigmatop = midintensity*(1+sigmarange);
double sigmabottom = midintensity*(1-sigmarange);
//calculate mean of values that differ are in sigma range.
for (ix=0;ix<kernel;ix++) {
for (iy=0;iy<kernel;iy++) {
v = ip.getPixelValue(x+ix-radius,y+iy-radius);
if ((v>=sigmabottom)&&(v<=sigmatop)){
sum = sum+v;
count = count+1; }
backupsum = v+ backupsum;
}
}
//if there are too few pixels in the kernal that are within sigma range, the
//mean of the entire kernal is taken. My modification of Lee's filter is to exclude the central value
//from the calculation of the mean as I assume it to be spuriously high or low
if (!(count>(minpixnumber)))
{sum = (backupsum-midintensity);
count = (int)((kernel*kernel)-1);
if (!modified)
{sum = (backupsum);
count = (int)(kernel*kernel);}
}
double val = (sum/count);
ip.putPixelValue(x,y, val);
n = n+1;
double percentage = (((double)n/status)*100);
IJ.showStatus(IJ.d2s(percentage,0) +"% done");
}
// IJ.showProgress(i, status);
}}
imp.updateAndDraw();
IJ.showStatus(IJ.d2s((System.currentTimeMillis()-start)/1000.0, 2)+" seconds"); }
private void geterrors() {
GenericDialog gd = new GenericDialog("Options");
float conf = 0.67f;
gd.addNumericField("Confidence Limit", (int) (conf * 100.0f), 5, 10, null);
gd.addChoice("Error Parameter", paramsnames, paramsnames[0]);
double spacing = 0.01;
gd.addNumericField("Chi^2 plot spacing (% of value)?", spacing * 100.0, 2, 10, null);
boolean globalerror = false;
gd.addCheckbox("Global Fit Error?", globalerror);
int dataset = 0;
gd.addNumericField("Data Set (for Global Error)", dataset, 0);
gd.showDialog();
if (gd.wasCanceled()) {
return;
}
conf = 0.01f * (float) gd.getNextNumber();
int paramindex = (int) gd.getNextChoiceIndex();
spacing = 0.01 * gd.getNextNumber();
globalerror = gd.getNextBoolean();
dataset = (int) gd.getNextNumber();
if (globalerror) {
support_plane_errors erclass = new support_plane_errors(this, 0.0001, 50, true, 0.1);
int[] erindeces = {paramindex, dataset};
// need to set up all the matrices
int nsel = 0;
int nparams = 11;
for (int i = 0; i < ncurves; i++) {
if (include[i]) {
nsel++;
}
}
double[][] params = new double[nsel][nparams];
String[][] tempformulas = new String[nsel][nparams];
double[][][] constraints = new double[2][nsel][nparams];
int[][] vflmatrix = new int[nsel][nparams];
float[][] tempdata = new float[nsel][xpts * ypts];
float[][] tempweights = new float[nsel][xpts * ypts];
int nfit = 0;
int counter = 0;
for (int i = 0; i < ncurves; i++) {
if (include[i]) {
for (int j = 0; j < nparams; j++) {
params[counter][j] = globalparams[i][j];
tempformulas[counter][j] = globalformulas[i][j];
constraints[0][counter][j] = globalconstraints[0][i][j];
constraints[1][counter][j] = globalconstraints[1][i][j];
vflmatrix[counter][j] = globalvflmatrix[i][j];
if (vflmatrix[counter][j] == 0 || (j == 0 && vflmatrix[counter][j] == 2)) {
nfit++;
}
}
for (int j = 0; j < xpts; j++) {
for (int k = 0; k < ypts; k++) {
tempdata[counter][j + k * xpts] = (float) ((double) pch[i][j][k] / (double) nmeas[i]);
tempweights[counter][j + k * xpts] = weights[i][j][k];
}
}
counter++;
}
}
int dofnum = xpts * ypts * nsel - (nfit - 1) - 1;
int dofden = xpts * ypts * nsel - nfit - 1;
// double flim=FLimit(dofnum,dofden,(double)conf);
double flim = (new jdist()).FLimit(dofnum, dofden, (double) conf);
IJ.log("FLimit = " + (float) flim);
if (flim == Double.NaN && flim < 1.0) {
IJ.showMessage("Invalid Limiting F Value");
return;
}
double truespacing = Math.abs(params[erindeces[1]][erindeces[0]] * spacing);
double[][] c2plot =
erclass.geterrorsglobal(
params,
vflmatrix,
tempformulas,
paramsnames,
constraints,
tempdata,
tempweights,
flim,
truespacing,
erindeces);
IJ.log("upper limit = " + c2plot[1][0] + " lower limit = " + c2plot[0][0]);
int templength = c2plot[0].length;
float[][] c2plotf = new float[2][templength - 1];
for (int i = 0; i < (templength - 1); i++) {
c2plotf[0][i] = (float) c2plot[0][i + 1];
c2plotf[1][i] = (float) c2plot[1][i + 1];
}
new PlotWindow4(
"c2 plot",
paramsnames[paramindex] + "[" + dataset + "]",
"Chi^2",
c2plotf[0],
c2plotf[1])
.draw();
} else {
support_plane_errors erclass = new support_plane_errors(this, 0.0001, 50, false, 0.1);
int errindex = paramindex;
float[] tempdata = new float[xpts * ypts];
float[] tempweights = new float[xpts * ypts];
for (int i = 0; i < xpts; i++) {
for (int j = 0; j < ypts; j++) {
tempdata[i + j * xpts] = (float) ((double) avg[i][j] / (double) nmeas[ncurves]);
tempweights[i + j * xpts] = avgweights[i][j];
}
}
int nfit = 0;
for (int i = 0; i < 7; i++) {
if (avgfixes[i] == 0) {
nfit++;
}
}
int dofnum = xpts * ypts - (nfit - 1) - 1;
int dofden = xpts * ypts - nfit - 1;
double flim = (new jdist()).FLimit(dofnum, dofden, (double) conf);
IJ.log("FLimit = " + (float) flim);
if (flim == Double.NaN && flim < 1.0) {
IJ.showMessage("Invalid Limiting F Value");
return;
}
double truespacing = Math.abs(avgparams[errindex] * spacing);
double[][] c2plot =
erclass.geterrors(
avgparams,
avgfixes,
avgconstraints,
tempdata,
tempweights,
flim,
truespacing,
errindex);
IJ.log("upper limit = " + c2plot[1][0] + " lower limit = " + c2plot[0][0]);
int templength = c2plot[0].length;
float[][] c2plotf = new float[2][templength - 1];
for (int i = 0; i < (templength - 1); i++) {
c2plotf[0][i] = (float) c2plot[0][i + 1];
c2plotf[1][i] = (float) c2plot[1][i + 1];
}
new PlotWindow4("c2 plot", paramsnames[errindex], "Chi^2", c2plotf[0], c2plotf[1]).draw();
}
}
public void run(String arg) {
GenericDialog gd = new GenericDialog("Options");
double sfreq = 20000.0;
gd.addNumericField("Sampling Frequency?", sfreq, 1, 10, null);
String[] psfchoice = {"3D Gaussian", "Gaus-Lorentz^2", "2D Gaussian"};
gd.addChoice("PSF Type?", psfchoice, psfchoice[0]);
String[] filetypechoice = {
"Confocor 3 raw", "Short binary trajectory", "PlotWindow trajectory", "Ascii Text File"
};
gd.addChoice("File Type?", filetypechoice, filetypechoice[0]);
boolean ch2green = true;
gd.addCheckbox("Ch2 is green?", ch2green);
gd.showDialog();
if (gd.wasCanceled()) {
return;
}
sfreq = gd.getNextNumber();
int psfflag = gd.getNextChoiceIndex();
int fileflag = gd.getNextChoiceIndex();
ch2green = gd.getNextBoolean();
int nfiles = 0;
Object[] histograms = null;
int xmax = 0;
int ymax = 0;
String[] names = null;
if (fileflag < 2) {
jdataio ioclass = new jdataio();
File[] filearray = ioclass.openfiles(OpenDialog.getDefaultDirectory(), IJ.getInstance());
if (filearray.length == 0) {
return;
}
String dir = filearray[0].getAbsolutePath();
int sepindex = dir.lastIndexOf(File.separator);
String newdir = dir.substring(0, sepindex + 1);
OpenDialog.setDefaultDirectory(newdir);
nfiles = filearray.length / 2;
if (nfiles > 25) {
nfiles = 25;
}
histograms = new Object[nfiles];
names = organize_c3_files(filearray);
for (int i = 0; i < nfiles; i++) {
try {
int length1 = (int) (((double) filearray[2 * i].length() - 128.0) / 4.0);
int length2 = (int) (((double) filearray[2 * i + 1].length() - 128.0) / 4.0);
int length3 = (int) (((double) filearray[2 * i].length()) / 2.0);
int length4 = (int) (((double) filearray[2 * i + 1].length()) / 2.0);
InputStream instream = new BufferedInputStream(new FileInputStream(filearray[2 * i]));
InputStream instream2 =
new BufferedInputStream(new FileInputStream(filearray[2 * i + 1]));
if (fileflag == 0) {
int[] pmdata = new int[length1];
int[] pmdata2 = new int[length2];
if (!ioclass.skipstreambytes(instream, 128)) {
showioerror();
instream.close();
return;
}
if (!ioclass.skipstreambytes(instream2, 128)) {
showioerror();
instream2.close();
return;
}
if (!ioclass.readintelintfile(instream, length1, pmdata)) {
showioerror();
instream.close();
return;
}
if (!ioclass.readintelintfile(instream2, length2, pmdata2)) {
showioerror();
instream2.close();
return;
}
if (ch2green) {
histograms[i] = (new pmodeconvert()).pm2pch(pmdata2, pmdata, sfreq, 20000000);
} else {
histograms[i] = (new pmodeconvert()).pm2pch(pmdata, pmdata2, sfreq, 20000000);
}
} else {
float[] tmdata = new float[length3];
float[] tmdata2 = new float[length4];
if (!ioclass.readintelshortfile(instream, length3, tmdata)) {
showioerror();
instream.close();
return;
}
if (!ioclass.readintelshortfile(instream2, length4, tmdata2)) {
showioerror();
instream2.close();
return;
}
if (ch2green) {
histograms[i] = (new pmodeconvert()).create_2Dhistogram(tmdata2, tmdata);
} else {
histograms[i] = (new pmodeconvert()).create_2Dhistogram(tmdata, tmdata2);
}
}
if (((float[][]) histograms[i]).length > xmax) {
xmax = ((float[][]) histograms[i]).length;
}
if (((float[][]) histograms[i])[0].length > ymax) {
ymax = ((float[][]) histograms[i])[0].length;
}
instream.close();
instream2.close();
} catch (IOException e) {
showioerror();
return;
}
}
} else {
if (fileflag == 2) {
ImageWindow iw = WindowManager.getCurrentWindow();
float[][] trajectories = (float[][]) jutils.runPW4VoidMethod(iw, "getYValues");
float[][] tempxvals = (float[][]) jutils.runPW4VoidMethod(iw, "getXValues");
sfreq = 1.0 / ((double) tempxvals[0][1]);
nfiles = trajectories.length / 2;
if (nfiles > 25) {
nfiles = 25;
}
names = new String[nfiles + 1];
names[nfiles] = "avg";
histograms = new Object[nfiles];
for (int i = 0; i < nfiles; i++) {
names[i] = "trajectory " + (i + 1);
if (ch2green) {
histograms[i] =
(new pmodeconvert())
.create_2Dhistogram(trajectories[2 * i + 1], trajectories[2 * i]);
} else {
histograms[i] =
(new pmodeconvert())
.create_2Dhistogram(trajectories[2 * i], trajectories[2 * i + 1]);
}
if (((float[][]) histograms[i]).length > xmax) {
xmax = ((float[][]) histograms[i]).length;
}
if (((float[][]) histograms[i])[0].length > ymax) {
ymax = ((float[][]) histograms[i])[0].length;
}
}
} else {
// here we read tab delimited lines from files
jdataio ioclass = new jdataio();
File[] filearray = ioclass.openfiles(OpenDialog.getDefaultDirectory(), IJ.getInstance());
if (filearray.length == 0) {
return;
}
String dir = filearray[0].getAbsolutePath();
int sepindex = dir.lastIndexOf(File.separator);
String newdir = dir.substring(0, sepindex + 1);
OpenDialog.setDefaultDirectory(newdir);
nfiles = filearray.length;
if (nfiles > 25) {
nfiles = 25;
}
histograms = new Object[nfiles];
names = new String[nfiles + 1];
names[nfiles] = "avg";
for (int i = 0; i < nfiles; i++) {
try {
names[i] = filearray[i].getName();
BufferedReader d = new BufferedReader(new FileReader(filearray[i]));
String[] lines = new String[256];
int counter = 0;
do {
lines[counter] = d.readLine();
counter++;
} while ((lines[counter - 1] != null && lines[counter - 1] != "") && counter < 256);
int numcolumns = 0;
for (int j = 0; j < counter - 1; j++) {
int temp = getncolumns(lines[j]);
if (temp > numcolumns) {
numcolumns = temp;
}
}
float[][] temphist2 = null;
if (ch2green) {
temphist2 = new float[numcolumns][counter - 1];
} else {
temphist2 = new float[counter - 1][numcolumns];
}
for (int k = 0; k < counter - 1; k++) {
float[] temp = tab_delim2float(lines[k]);
for (int j = 0; j < numcolumns; j++) {
if (ch2green) {
temphist2[j][k] = temp[j];
} else {
temphist2[k][j] = temp[j];
}
}
}
histograms[i] = temphist2;
d.close();
} catch (IOException e) {
showioerror();
return;
}
}
for (int i = 0; i < nfiles; i++) {
if (((float[][]) histograms[i]).length > xmax) {
xmax = ((float[][]) histograms[i]).length;
}
if (((float[][]) histograms[i])[0].length > ymax) {
ymax = ((float[][]) histograms[i])[0].length;
}
}
}
}
// note that here x is green and y is red
float[][][] pch = new float[nfiles][xmax][ymax];
for (int i = 0; i < nfiles; i++) {
for (int j = 0; j < ((float[][]) histograms[i]).length; j++) {
for (int k = 0; k < ((float[][]) histograms[i])[j].length; k++) {
pch[i][j][k] = ((float[][]) histograms[i])[j][k];
}
}
}
final PCH2DFitWindow cw = new PCH2DFitWindow();
cw.init(names, pch, psfflag);
final Frame f = new Frame("PCH 2D Analysis");
f.setLocation(10, 10);
f.addWindowListener(
new WindowAdapter() {
public void windowClosing(WindowEvent e) {
f.dispose();
}
});
f.add(cw);
f.pack();
f.setResizable(false);
Insets ins = f.getInsets();
cw.totalSize.height = PCH2DFitWindow.H + ins.bottom + ins.top + 65;
cw.totalSize.width = PCH2DFitWindow.WR + ins.left + ins.right;
f.setSize(cw.totalSize);
f.setVisible(true);
cw.requestFocus();
}
public boolean showDialog() {
String bgChoice = "NoBg";
int[] wList = WindowManager.getIDList();
if (wList == null) {
IJ.noImage();
return false;
}
String[] sampleTitles = new String[wList.length];
for (int i = 0; i < wList.length; i++) {
ImagePlus imp = WindowManager.getImage(wList[i]);
sampleTitles[i] = imp != null ? imp.getTitle() : "";
}
String[] bgTitles = new String[wList.length + 1];
bgTitles[0] = "NoBg";
for (int i = 1; i < wList.length + 1; i++) {
bgTitles[i] = sampleTitles[i - 1];
}
for (int i = 1; i < (wList.length + 1); i++) {
if (bgTitles[i] == bgStackTitle) bgChoice = bgStackTitle;
}
String sampleChoice = sampleTitles[0];
for (int i = 1; i < (wList.length); i++) {
if (sampleTitles[i] == sampleStackTitle) sampleChoice = sampleStackTitle;
}
String[] mirrors = new String[2];
mirrors[0] = "No";
mirrors[1] = "Yes";
GenericDialog gd = new GenericDialog("PolScope 5Frame Calc");
gd.addChoice("Sample:", sampleTitles, sampleChoice);
gd.addChoice("Background:", bgTitles, bgChoice);
gd.addChoice("Mirror:", mirrors, mirror);
gd.addNumericField("Wavelength: ", wavelength, 1, 8, " nm");
gd.addNumericField("Swing: ", swing, 3, 8, " wavelength");
gd.addNumericField("Ret. Ceiling: ", retCeiling, 1, 8, " nm");
gd.addNumericField("Orient. Ref.: ", azimRef, 1, 8, " degree");
gd.showDialog();
if (gd.wasCanceled()) return false;
int index1 = gd.getNextChoiceIndex();
int index2 = gd.getNextChoiceIndex();
int index3 = gd.getNextChoiceIndex();
wavelength = (float) gd.getNextNumber();
swing = (float) gd.getNextNumber();
retCeiling = (float) gd.getNextNumber();
azimRef = (float) gd.getNextNumber();
imp1 = WindowManager.getImage(wList[index1]);
sampleStackTitle = sampleTitles[index1];
bgStackTitle = bgTitles[index2];
if (bgStackTitle == "NoBg") {
imp2 =
WindowManager.getImage(
wList[
index1]); // only to assign a valid ImagePlus to imp2 which is not used when NoBg
} else {
imp2 = WindowManager.getImage(wList[index2 - 1]);
}
mirror = mirrors[index3];
return true;
}
public void run(String arg) {
String[] labels = {
"Masked_Chromosomes", "Unmixed_Image", "Spectral_Image(optional)", "Spectra(optional)"
};
ImagePlus[] imps = jutils.selectImages(true, 4, labels);
if (imps == null) {
return;
}
if (imps[0] == null) {
return;
}
float[] mask = (float[]) imps[0].getStack().getPixels(2);
findblobs3 fb = new findblobs3(imps[0].getWidth(), imps[0].getHeight());
float[] objects = fb.dofindblobs(mask, 0.5f);
WaitForUserDialog dg =
new WaitForUserDialog(
"Optional Input", "Place RoiManager Points on Chromosome Segments (if desired)");
dg.show();
if (!dg.escPressed()) {
RoiManager rman = RoiManager.getInstance();
while (rman != null && rman.getCount() > 1) {
Roi[] rois = rman.getRoisAsArray();
int[] ids = new int[rois.length];
for (int i = 0; i < rois.length; i++) {
Rectangle r = rois[i].getBounds();
ids[i] = (int) objects[r.x + fb.width * r.y];
}
objects = fb.link_objects(objects, ids);
rman.reset();
dg =
new WaitForUserDialog(
"Optional Input",
"Place More RoiManager Points on Chromosome Segments (if desired)");
dg.show();
if (dg.escPressed()) break;
}
}
int[] areas = fb.get_areas(objects);
int[] temprank = jsort.get_javasort_order(areas);
int[] arearank = jsort.get_javasort_order(temprank);
for (int i = 0; i < fb.nobjects; i++) {
arearank[i] = fb.nobjects - arearank[i] - 1;
}
// if the spectra are available, get them
float[][][] spectra = null;
Object[] data = null;
if (imps[1] != null && imps[2] != null && imps[3] != null) {
ImageWindow iw = imps[3].getWindow();
if (iw.getClass().getName().equals("jguis.PlotWindow4")) {
float[][] yvals = (float[][]) jutils.runPW4VoidMethod(iw, "getYValues");
data = jutils.stack2array(imps[2].getStack());
Object[] coef = jutils.stack2array(imps[1].getStack());
spectra = new float[fb.nobjects][2][];
for (int i = 0; i < fb.nobjects; i++) {
spectra[i][0] = fb.get_object_spectrum(objects, (i + 1), data, "Sum");
spectra[i][1] = new float[yvals[0].length];
float[] tempcoef = fb.get_object_spectrum(objects, (i + 1), coef, "Sum");
for (int j = 0; j < yvals[0].length; j++) {
for (int k = 0; k < 5; k++) {
spectra[i][1][j] += tempcoef[k] * yvals[k][j];
}
}
}
}
}
CompositeImage imp = (CompositeImage) imps[0];
imp.setPosition(1, 1, 1);
LUT graylut = jutils.get_lut_for_color(Color.white);
imp.setChannelColorModel(graylut);
imp.setPosition(2, 1, 1);
LUT redlut = jutils.get_lut_for_color(Color.red);
imp.setChannelColorModel(redlut);
imp.setPosition(1, 1, 1);
imp.updateAndRepaintWindow();
SkyPanel_v3 sp = new SkyPanel_v3();
int skychan = 6;
if (imps[1] != null) skychan = imps[1].getNChannels();
// assume that the sky image has 6 channels and that the second is the unknown green
// shift the unknown green to the end
ImagePlus skyimp = null;
if (imps[1] != null) {
Object[] skystack = jutils.stack2array(imps[1].getStack());
// Object[]
// skystack2={skystack[0],skystack[2],skystack[3],skystack[4],skystack[5],skystack[1]};
Object[] skystack2 = null;
if (skychan == 6)
skystack2 = new Object[] {skystack[0], skystack[2], skystack[3], skystack[4], skystack[5]};
else
skystack2 = new Object[] {skystack[0], skystack[1], skystack[2], skystack[3], skystack[4]};
skyimp =
new ImagePlus(
"rearranged", jutils.array2stack(skystack2, imps[1].getWidth(), imps[1].getHeight()));
}
int nch = 5;
if (skyimp != null) nch = skyimp.getStack().getSize();
GenericDialog gd2 = new GenericDialog("Options");
gd2.addNumericField("Area Accuracy (percent)", 30, 0);
for (int i = 0; i < nch; i++) {
gd2.addNumericField("Ch_" + (i + 1) + "_Contr_Thresh", 0.35, 5, 15, null);
}
// gd2.addNumericField("Contribution Threshold",0.35,5,15,null);
gd2.addCheckbox("Mouse?", false);
gd2.addNumericField("Box_Width", 150, 0);
gd2.addNumericField("Box_Height", 100, 0);
gd2.showDialog();
if (gd2.wasCanceled()) {
return;
}
sp.areathresh = (float) gd2.getNextNumber();
sp.objthresh2 = new float[nch];
for (int i = 0; i < nch; i++) sp.objthresh2[i] = (float) gd2.getNextNumber();
// sp.objthresh=(float)gd2.getNextNumber();
boolean mouse = gd2.getNextBoolean();
int bwidth = (int) gd2.getNextNumber();
int bheight = (int) gd2.getNextNumber();
int[] colorindices = {4, 1, 2, 6, 3};
GenericDialog gd3 = new GenericDialog("Color Options");
for (int i = 0; i < 5; i++)
gd3.addChoice(
"Ch" + (i + 1) + " Color",
SkyPanel_v3.colornames,
SkyPanel_v3.colornames[colorindices[i]]);
gd3.showDialog();
if (gd3.wasCanceled()) return;
for (int i = 0; i < 5; i++) colorindices[i] = gd3.getNextChoiceIndex();
sp.colorindices = colorindices;
sp.nch = 5;
sp.dapilast = false;
sp.cellwidth = bwidth;
sp.cellheight = bheight;
sp.init(imps[0], skyimp, objects, areas, arearank, fb, true, spectra, data, mouse);
SkyPanel_v3.launch_frame(sp);
}
public boolean get_errors(double[] params, int[] fixes) {
GenericDialog gd = new GenericDialog("Error Options");
String[] methods = {"Support Plane", "Monte Carlo"};
gd.addChoice("Method", methods, methods[0]);
float conf = 0.67f;
gd.addNumericField("SP_Confidence Limit (%)", (int) (conf * 100.0f), 5, 10, null);
String[] labels = {"P1", "P2", "P3", "P4", "P5", "P6", "P7", "P8", "P9", "P10"};
gd.addChoice("SP_Parameter", labels, labels[0]);
double spacing = 0.01;
gd.addNumericField("SP_Chi^2_plot_spacing (% of value)?", spacing * 100.0, 2, 10, null);
int ntrials = 100;
gd.addNumericField("MC_#_Trials", ntrials, 0);
gd.showDialog();
if (gd.wasCanceled()) {
return false;
}
int methodindex = gd.getNextChoiceIndex();
conf = 0.01f * (float) gd.getNextNumber();
int paramindex = gd.getNextChoiceIndex();
spacing = 0.01 * gd.getNextNumber();
ntrials = (int) gd.getNextNumber();
if (methodindex == 0) {
support_plane_errors_v2 erclass = new support_plane_errors_v2(this, 0.0001, 50, false, 0.1);
int errindex = paramindex;
int nfit = 0;
for (int i = 0; i < labels.length; i++) {
if (fixes[i] == 0) {
nfit++;
}
}
int npts = tempdata.length;
int dofnum = npts - (nfit - 1) - 1;
int dofden = npts - nfit - 1;
double flim = (new jdist()).FLimit(dofnum, dofden, (double) conf);
IJ.log("FLimit = " + (float) flim);
if (flim == Double.NaN && flim < 1.0) {
IJ.showMessage("Invalid Limiting F Value");
return false;
}
double truespacing = Math.abs(params[errindex] * spacing);
double[][] c2plot =
erclass.geterrors(
params, fixes, constraints, tempdata, weights, flim, truespacing, errindex);
IJ.log("upper limit = " + c2plot[1][0] + " lower limit = " + c2plot[0][0]);
IJ.log(
"upper error = "
+ (c2plot[1][0] - params[errindex])
+ " lower error = "
+ (params[errindex] - c2plot[0][0]));
int templength = c2plot[0].length;
float[][] c2plotf = new float[2][templength - 1];
for (int i = 0; i < (templength - 1); i++) {
c2plotf[0][i] = (float) c2plot[0][i + 1];
c2plotf[1][i] = (float) c2plot[1][i + 1];
}
new PlotWindow4("c2 plot", labels[errindex], "Chi^2", c2plotf[0], c2plotf[1]).draw();
} else {
StringBuffer sb = new StringBuffer();
sb.append("Trial\t");
for (int i = 0; i < labels.length; i++) {
if (fixes[i] == 0) sb.append(labels[i] + "\t");
}
sb.append("chi^2");
tw = new TextWindow("Monte Carlo Results", sb.toString(), "", 400, 400);
redirect = true;
monte_carlo_errors_v2 erclass = new monte_carlo_errors_v2(this, 0.0001, 50, false, 0.1);
double[][] errors = erclass.geterrors(params, fixes, constraints, tempdata, weights, ntrials);
sb = new StringBuffer();
sb.append("StDev\t");
for (int i = 0; i < errors.length; i++) {
float[] ferr = new float[errors[0].length];
for (int j = 0; j < ferr.length; j++) ferr[j] = (float) errors[i][j];
float stdev = jstatistics.getstatistic("StDev", ferr, null);
sb.append("" + stdev);
if (i < (errors.length - 1)) sb.append("\t");
}
tw.append(sb.toString());
redirect = false;
}
return true;
}
/** Ask for parameters and then execute. */
public void run(String arg) {
// 1 - Obtain the currently active image:
ImagePlus imp = IJ.getImage();
if (null == imp) {
IJ.showMessage("There must be at least one image open");
return;
}
if (imp.getBitDepth() != 8) {
IJ.showMessage("Error", "Only 8-bit images are supported");
return;
}
// 2 - Ask for parameters:
GenericDialog gd = new GenericDialog("Auto Local Threshold");
String[] methods = {
"Try all",
"Bernsen",
"Contrast",
"Mean",
"Median",
"MidGrey",
"Niblack",
"Otsu",
"Phansalkar",
"Sauvola"
};
gd.addMessage("Auto Local Threshold v1.5");
gd.addChoice("Method", methods, methods[0]);
gd.addNumericField("Radius", 15, 0);
gd.addMessage("Special paramters (if different from default)");
gd.addNumericField("Parameter_1", 0, 0);
gd.addNumericField("Parameter_2", 0, 0);
gd.addCheckbox("White objects on black background", true);
if (imp.getStackSize() > 1) {
gd.addCheckbox("Stack", false);
}
gd.addMessage("Thresholded result is always shown in white [255].");
gd.showDialog();
if (gd.wasCanceled()) return;
// 3 - Retrieve parameters from the dialog
String myMethod = gd.getNextChoice();
int radius = (int) gd.getNextNumber();
double par1 = (double) gd.getNextNumber();
double par2 = (double) gd.getNextNumber();
boolean doIwhite = gd.getNextBoolean();
boolean doIstack = false;
int stackSize = imp.getStackSize();
if (stackSize > 1) doIstack = gd.getNextBoolean();
// 4 - Execute!
// long start = System.currentTimeMillis();
if (myMethod.equals("Try all")) {
ImageProcessor ip = imp.getProcessor();
int xe = ip.getWidth();
int ye = ip.getHeight();
int ml = methods.length;
ImagePlus imp2, imp3;
ImageStack tstack = null, stackNew;
if (stackSize > 1 && doIstack) {
boolean doItAnyway = true;
if (stackSize > 25) {
YesNoCancelDialog d =
new YesNoCancelDialog(
IJ.getInstance(),
"Auto Local Threshold",
"You might run out of memory.\n \nDisplay "
+ stackSize
+ " slices?\n \n \'No\' will process without display and\noutput results to the log window.");
if (!d.yesPressed()) {
// doIlog=true; //will show in the log window
doItAnyway = false;
}
if (d.cancelPressed()) return;
}
for (int j = 1; j <= stackSize; j++) {
imp.setSlice(j);
ip = imp.getProcessor();
tstack = new ImageStack(xe, ye);
for (int k = 1; k < ml; k++) tstack.addSlice(methods[k], ip.duplicate());
imp2 = new ImagePlus("Auto Threshold", tstack);
imp2.updateAndDraw();
for (int k = 1; k < ml; k++) {
imp2.setSlice(k);
Object[] result = exec(imp2, methods[k], radius, par1, par2, doIwhite);
}
// if (doItAnyway){
CanvasResizer cr = new CanvasResizer();
stackNew = cr.expandStack(tstack, (xe + 2), (ye + 18), 1, 1);
imp3 = new ImagePlus("Auto Threshold", stackNew);
imp3.updateAndDraw();
MontageMaker mm = new MontageMaker();
mm.makeMontage(imp3, 3, 3, 1.0, 1, (ml - 1), 1, 0, true); // 3 columns and 3 rows
}
imp.setSlice(1);
// if (doItAnyway)
IJ.run("Images to Stack", "method=[Copy (center)] title=Montage");
return;
} else { // single image try all
tstack = new ImageStack(xe, ye);
for (int k = 1; k < ml; k++) tstack.addSlice(methods[k], ip.duplicate());
imp2 = new ImagePlus("Auto Threshold", tstack);
imp2.updateAndDraw();
for (int k = 1; k < ml; k++) {
imp2.setSlice(k);
// IJ.log("analyzing slice with "+methods[k]);
Object[] result = exec(imp2, methods[k], radius, par1, par2, doIwhite);
}
// imp2.setSlice(1);
CanvasResizer cr = new CanvasResizer();
stackNew = cr.expandStack(tstack, (xe + 2), (ye + 18), 1, 1);
imp3 = new ImagePlus("Auto Threshold", stackNew);
imp3.updateAndDraw();
MontageMaker mm = new MontageMaker();
mm.makeMontage(imp3, 3, 3, 1.0, 1, (ml - 1), 1, 0, true);
return;
}
} else { // selected a method
if (stackSize > 1 && doIstack) { // whole stack
// if (doIstackHistogram) {// one global histogram
// Object[] result = exec(imp, myMethod, noWhite, noBlack, doIwhite, doIset, doIlog,
// doIstackHistogram );
// }
// else{ // slice by slice
for (int k = 1; k <= stackSize; k++) {
imp.setSlice(k);
Object[] result = exec(imp, myMethod, radius, par1, par2, doIwhite);
}
// }
imp.setSlice(1);
} else { // just one slice
Object[] result = exec(imp, myMethod, radius, par1, par2, doIwhite);
}
// 5 - If all went well, show the image:
// not needed here as the source image is binarised
}
}
public boolean reconstructDataset() {
view_modes[0] = "3D color";
view_modes[1] = "2D histogram";
view_modes[2] = "2D particle intensity (16-bit)";
view_modes[3] = "2D particle intensity (8-bit)";
GenericDialog gd = new GenericDialog("Reconstruct PALM/STORM Dataset");
gd.addNumericField(
"Target pixel size for the rendered image (nm)",
prefs.get("QuickPALM.viewer_tpixelsize", 30),
2);
gd.addNumericField("Original image width (px)", prefs.get("QuickPALM.viewer_owidth", 512), 2);
gd.addNumericField("Original image height (px)", prefs.get("QuickPALM.viewer_oheight", 512), 2);
gd.addChoice("View mode", view_modes, prefs.get("QuickPALM.view_mode", view_modes[1]));
// gd.addNumericField("Allow image saturation (%)", prefs.get("QuickPALM.saturation", 50), 0);
gd.addCheckbox(
"Simulate sub-difraction spot (gaussian convolution - only 2D)",
prefs.get("QuickPALM.viewer_doConvolve", true));
// gd.addCheckbox("Make 3D stack", prefs.get("QuickPALM.viewer_do3d", false));
// gd.addCheckbox("Make movie", prefs.get("QuickPALM.viewer_doMovie", false));
gd.addCheckbox("Make 3D stack", false);
gd.addCheckbox("Make movie", false);
// gd.addCheckbox("Save only and don't show", prefs.get("QuickPALM.viewer_doSave", false));
gd.addMessage("\n");
// -----------------------------------------
gd.addMessage("-- Simulate sub-difraction spot settings (used only if selected) --");
gd.addNumericField("FWHM of the spot", prefs.get("QuickPALM.viewer_fwhm", 30), 2);
gd.addMessage("\n");
// -----------------------------------------
gd.addMessage("-- Make 3D stack settings (used only if selected) --");
gd.addNumericField("Z-spacing between slices (nm)", prefs.get("QuickPALM.viewer_zstep", 50), 2);
gd.addNumericField(
"Merge particle Z-position above (nm - 0 for full Z range)",
prefs.get("QuickPALM.viewer_mergeabove", 0),
2);
gd.addNumericField(
"Merge particle Z-position bellow (nm - 0 for full Z range)",
prefs.get("QuickPALM.viewer_mergebellow", 0),
2);
gd.addMessage("\n");
// -----------------------------------------
gd.addMessage("-- Make movie settings (used only if selected) --");
gd.addNumericField(
"Make a reconstruction in every N frames", prefs.get("QuickPALM.viewer_update", 10), 0);
gd.addNumericField(
"Accumulate N neighboring frames for each reconstruction\n(set to 0 to accumulate all the preceding frames)",
prefs.get("QuickPALM.viewer_accumulate", 100),
0);
gd.showDialog();
if (gd.wasCanceled()) return false;
viewer_tpixelsize = gd.getNextNumber();
prefs.set("QuickPALM.viewer_tpixelsize", viewer_tpixelsize);
viewer_owidth = (int) gd.getNextNumber();
prefs.set("QuickPALM.viewer_owidth", viewer_owidth);
viewer_oheight = (int) gd.getNextNumber();
prefs.set("QuickPALM.viewer_oheight", viewer_oheight);
view_mode = gd.getNextChoice();
prefs.set("QuickPALM.view_mode", view_mode);
viewer_doConvolve = gd.getNextBoolean();
prefs.set("QuickPALM.viewer_doConvolve", viewer_doConvolve);
viewer_do3d = gd.getNextBoolean();
prefs.set("QuickPALM.viewer_do3d", viewer_do3d);
viewer_doMovie = gd.getNextBoolean();
prefs.set("QuickPALM.viewer_doMovie", viewer_doMovie);
// viewer_doSave = gd.getNextBoolean();
// prefs.set("QuickPALM.viewer_doSave", viewer_doSave);
// -- Simulate sub-difraction spot
viewer_fwhm = gd.getNextNumber();
prefs.set("QuickPALM.viewer_fwhm", viewer_fwhm);
// -- Show B&W
// viewer_is8bit = gd.getNextBoolean();
// prefs.set("QuickPALM.viewer_is8bit", viewer_is8bit);
// -- Make 3D stack
viewer_zstep = gd.getNextNumber();
prefs.set("QuickPALM.viewer_zstep", viewer_zstep);
viewer_mergeabove = gd.getNextNumber();
prefs.set("QuickPALM.viewer_mergeabove", viewer_mergeabove);
viewer_mergebellow = gd.getNextNumber();
prefs.set("QuickPALM.viewer_mergebellow", viewer_mergebellow);
// -- Make Movie
viewer_update = (int) gd.getNextNumber();
prefs.set("QuickPALM.viewer_update", viewer_update);
viewer_accumulate = (int) gd.getNextNumber();
prefs.set("QuickPALM.viewer_accumulate", viewer_accumulate);
return true;
}
public void run(String arg) {
ImagePlus imp = WindowManager.getCurrentImage();
Calibration cal = imp.getCalibration();
GenericDialog gd = new GenericDialog("Options");
int subsize = 32;
gd.addNumericField("Subregion Size (pixels)?", subsize, 0);
int stepsize = 16;
gd.addNumericField("Step Size?", stepsize, 0);
int shift = 3;
gd.addNumericField("STICS temporal Shift?", shift, 0);
float xoffset = 0.0f;
gd.addNumericField("X_Offset", xoffset, 5, 15, null);
float yoffset = 0.0f;
gd.addNumericField("Y_Offset", yoffset, 5, 15, null);
float multiplier = 8.0f;
gd.addNumericField("Velocity Multiplier", multiplier, 5, 15, null);
float ftime = 1.0f;
gd.addNumericField("Frame_Time(min)", ftime, 5, 15, null);
float scaling = (float) cal.pixelWidth;
gd.addNumericField("Pixel_Size(um)", scaling, 5, 15, null);
boolean norm = true;
gd.addCheckbox("Normalize_Vector_lengths?", norm);
boolean centered = true;
gd.addCheckbox("Center_Vectors?", centered);
float magthresh = 0.0f;
gd.addNumericField("Magnitude_Threshhold?", magthresh, 5, 15, null);
int rlength = 10;
gd.addNumericField("Running_avg_length", rlength, 0);
int inc = 5;
gd.addNumericField("Start_frame_increment", inc, 0);
gd.showDialog();
if (gd.wasCanceled()) {
return;
}
subsize = (int) gd.getNextNumber();
stepsize = (int) gd.getNextNumber();
shift = (int) gd.getNextNumber();
xoffset = (float) gd.getNextNumber();
yoffset = (float) gd.getNextNumber();
multiplier = (float) gd.getNextNumber();
ftime = (float) gd.getNextNumber();
scaling = (float) gd.getNextNumber();
norm = gd.getNextBoolean();
centered = gd.getNextBoolean();
magthresh = (float) gd.getNextNumber();
rlength = (int) gd.getNextNumber();
inc = (int) gd.getNextNumber();
int width = imp.getWidth();
int xregions = 1 + (int) (((float) width - (float) subsize) / (float) stepsize);
int newwidth = xregions * subsize;
int height = imp.getHeight();
int yregions = 1 + (int) (((float) height - (float) subsize) / (float) stepsize);
int newheight = yregions * subsize;
ImageStack stack = imp.getStack();
int slices = imp.getNSlices();
int channels = imp.getNChannels();
int frames = imp.getNFrames();
if (frames == 1) {
frames = slices;
slices = 1;
}
Roi roi = imp.getRoi();
if (roi == null) {
roi = new Roi(0, 0, width, height);
}
STICS_map map = new STICS_map(subsize, stepsize);
Object[] tseries = jutils.get3DTSeries(stack, 0, 0, frames, slices, channels);
map.update_STICS_map(tseries, width, height, 0, rlength, roi.getPolygon(), shift);
FloatProcessor fp =
map.get_map(scaling, ftime, stepsize, centered, norm, multiplier, stepsize, magthresh);
ImageStack vector_stack = new ImageStack(fp.getWidth(), fp.getHeight());
vector_stack.addSlice("", fp);
float[][] vel = map.get_scaled_velocities(scaling, ftime, stepsize);
ImageStack velstack = new ImageStack(map.xregions, map.yregions);
velstack.addSlice("", vel[0]);
velstack.addSlice("", vel[1]);
int velframes = 2;
IJ.showStatus("frame " + 0 + " calculated");
for (int i = inc; i < (frames - rlength); i += inc) {
map.update_STICS_map(tseries, width, height, i, rlength, roi.getPolygon(), shift);
FloatProcessor fp2 =
map.get_map(scaling, ftime, stepsize, centered, norm, multiplier, stepsize, magthresh);
vector_stack.addSlice("", fp2);
vel = map.get_scaled_velocities(scaling, ftime, stepsize);
velstack.addSlice("", vel[0]);
velstack.addSlice("", vel[1]);
velframes += 2;
IJ.showStatus("frame " + i + " calculated");
}
(new ImagePlus("STICS Vectors", vector_stack)).show();
ImagePlus imp3 = new ImagePlus("Velocities", velstack);
imp3.setOpenAsHyperStack(true);
imp3.setDimensions(2, 1, velframes / 2);
new CompositeImage(imp3, CompositeImage.COLOR).show();
}
public boolean analyseParticles(MyFunctions f) {
GenericDialog gd = new GenericDialog("Analyse PALM/STORM Particles");
gd.addNumericField("Minimum SNR", prefs.get("QuickPALM.snr", 5), 2);
gd.addNumericField("Maximum FWHM (in px)", prefs.get("QuickPALM.fwhm", 4), 0);
gd.addNumericField("Image plane pixel size (nm)", prefs.get("QuickPALM.pixelsize", 106), 2);
gd.addCheckbox("Smart SNR", prefs.get("QuickPALM.smartsnr", true));
gd.addCheckbox(
"3D PALM (astigmatism) - will require calibration file",
prefs.get("QuickPALM.is3d", false));
gd.addCheckbox("Online rendering", prefs.get("QuickPALM.view", true));
gd.addCheckbox("Attach to running acquisition", prefs.get("QuickPALM.attach", false));
gd.addCheckbox("Stream particle info directly into file", prefs.get("QuickPALM.stream", true));
gd.addMessage("\n");
// -----------------------------------------
gd.addMessage("-- Online rendering settings (used only if selected) --");
gd.addMessage("\n");
gd.addNumericField("Pixel size of rendered image (nm)", 30, 2);
gd.addNumericField("Accumulate last (0 to accumulate all frames)", 0, 0);
gd.addNumericField("Update every (frames)", 10, 0);
// gd.addNumericField("Allow color saturation (%)", 50, 0);
gd.addMessage("\n");
// -----------------------------------------
gd.addMessage("-- Attach to running acquisition settings (used only if selected) --");
gd.addMessage("\n");
gd.addStringField(
"_Image name pattern (NN...NN represents the numerical change)",
prefs.get("QuickPALM.pattern", "imgNNNNNNNNN.tif"),
20);
gd.addNumericField("Start NN...NN with", 0, 0);
gd.addNumericField("In acquisition max. wait time for new image (ms)", 50, 0);
gd.addMessage("\n");
// -----------------------------------------
gd.addMessage("-- Advanced settings (don't normally need to be changed) --");
gd.addMessage("\n");
gd.addNumericField("_Minimum symmetry (%)", prefs.get("QuickPALM.symmetry", 50), 0);
gd.addNumericField(
"Local threshold (% maximum intensity)", prefs.get("QuickPALM.lthreshold", 20), 0);
gd.addNumericField("_Maximum iterations per frame", prefs.get("QuickPALM.maxiter", 1000), 0);
gd.addNumericField(
"Threads (each takes ~3*[frame size] in memory)", prefs.get("QuickPALM.nthreads", 50), 0);
gd.addMessage("\n\nDon't forget to save the table in the end...");
gd.showDialog();
if (gd.wasCanceled()) return false;
snr = (int) gd.getNextNumber();
prefs.set("QuickPALM.snr", snr);
fwhm = gd.getNextNumber();
prefs.set("QuickPALM.fwhm", fwhm);
pixelsize = gd.getNextNumber();
prefs.set("QuickPALM.pixelsize", pixelsize);
smartsnr = gd.getNextBoolean();
prefs.set("QuickPALM.smartsnr", smartsnr);
is3d = gd.getNextBoolean();
prefs.set("QuickPALM.is3d", is3d);
view = gd.getNextBoolean();
prefs.set("QuickPALM.view", view);
attach = gd.getNextBoolean();
prefs.set("QuickPALM.attach", attach);
if (gd.getNextBoolean()) {
f.psave = new ParticleSaver();
f.psave.setup();
prefs.set("QuickPALM.stream", true);
} else prefs.set("QuickPALM.stream", false);
// --
magn = pixelsize / gd.getNextNumber();
viewer_accumulate = (int) gd.getNextNumber();
viewer_update = (int) gd.getNextNumber();
// --
pattern = gd.getNextString().trim();
prefs.set("QuickPALM.pattern", pattern);
prefix = pattern.substring(0, pattern.indexOf("N"));
sufix = pattern.substring(pattern.lastIndexOf("N") + 1, pattern.length());
nimchars = pattern.split("N").length - 1;
nimstart = (int) gd.getNextNumber();
waittime = (int) gd.getNextNumber();
// --
symmetry = gd.getNextNumber() / 100;
prefs.set("QuickPALM.symmetry", symmetry);
pthrsh = gd.getNextNumber() / 100;
prefs.set("QuickPALM.lthreshold", pthrsh * 100);
maxpart = (int) gd.getNextNumber();
prefs.set("QuickPALM.maxiter", maxpart);
threads = (int) gd.getNextNumber();
prefs.set("QuickPALM.nthreads", threads);
return true;
}