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;
}
void interpolate() {
Roi roi = imp.getRoi();
if (roi == null) {
noRoi("Interpolate");
return;
}
if (roi.getType() == Roi.POINT) return;
if (IJ.isMacro() && Macro.getOptions() == null) Macro.setOptions("interval=1");
GenericDialog gd = new GenericDialog("Interpolate");
gd.addNumericField("Interval:", 1.0, 1, 4, "pixel");
gd.addCheckbox("Smooth", IJ.isMacro() ? false : smooth);
gd.showDialog();
if (gd.wasCanceled()) return;
double interval = gd.getNextNumber();
smooth = gd.getNextBoolean();
Undo.setup(Undo.ROI, imp);
FloatPolygon poly = roi.getInterpolatedPolygon(interval, smooth);
int t = roi.getType();
int type = roi.isLine() ? Roi.FREELINE : Roi.FREEROI;
if (t == Roi.POLYGON && interval > 1.0) type = Roi.POLYGON;
if ((t == Roi.RECTANGLE || t == Roi.OVAL || t == Roi.FREEROI) && interval >= 5.0)
type = Roi.POLYGON;
if ((t == Roi.LINE || t == Roi.FREELINE) && interval >= 5.0) type = Roi.POLYLINE;
if (t == Roi.POLYLINE && interval >= 1.0) type = Roi.POLYLINE;
ImageCanvas ic = imp.getCanvas();
if (poly.npoints <= 150 && ic != null && ic.getMagnification() >= 12.0)
type = roi.isLine() ? Roi.POLYLINE : Roi.POLYGON;
Roi p = new PolygonRoi(poly, type);
if (roi.getStroke() != null) p.setStrokeWidth(roi.getStrokeWidth());
p.setStrokeColor(roi.getStrokeColor());
p.setName(roi.getName());
transferProperties(roi, p);
imp.setRoi(p);
}
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;
}
private void makeBand(ImagePlus imp) {
Roi roi = imp.getRoi();
if (roi == null) {
noRoi("Make Band");
return;
}
if (!roi.isArea()) {
IJ.error("Make Band", "Area selection required");
return;
}
Calibration cal = imp.getCalibration();
double pixels = bandSize;
double size = pixels * cal.pixelWidth;
int decimalPlaces = 0;
if ((int) size != size) decimalPlaces = 2;
GenericDialog gd = new GenericDialog("Make Band");
gd.addNumericField("Band Size:", size, decimalPlaces, 4, cal.getUnits());
gd.showDialog();
if (gd.wasCanceled()) return;
size = gd.getNextNumber();
if (Double.isNaN(size)) {
IJ.error("Make Band", "invalid number");
return;
}
int n = (int) Math.round(size / cal.pixelWidth);
if (n > 255) {
IJ.error("Make Band", "Cannot make bands wider that 255 pixels");
return;
}
int width = imp.getWidth();
int height = imp.getHeight();
Rectangle r = roi.getBounds();
ImageProcessor ip = roi.getMask();
if (ip == null) {
ip = new ByteProcessor(r.width, r.height);
ip.invert();
}
ImageProcessor mask = new ByteProcessor(width, height);
mask.insert(ip, r.x, r.y);
ImagePlus edm = new ImagePlus("mask", mask);
boolean saveBlackBackground = Prefs.blackBackground;
Prefs.blackBackground = false;
IJ.run(edm, "Distance Map", "");
Prefs.blackBackground = saveBlackBackground;
ip = edm.getProcessor();
ip.setThreshold(0, n, ImageProcessor.NO_LUT_UPDATE);
int xx = -1, yy = -1;
for (int x = r.x; x < r.x + r.width; x++) {
for (int y = r.y; y < r.y + r.height; y++) {
if (ip.getPixel(x, y) < n) {
xx = x;
yy = y;
break;
}
}
if (xx >= 0 || yy >= 0) break;
}
int count = IJ.doWand(edm, xx, yy, 0, null);
if (count <= 0) {
IJ.error("Make Band", "Unable to make band");
return;
}
ShapeRoi roi2 = new ShapeRoi(edm.getRoi());
if (!(roi instanceof ShapeRoi)) roi = new ShapeRoi(roi);
ShapeRoi roi1 = (ShapeRoi) roi;
roi2 = roi2.not(roi1);
imp.setRoi(roi2);
bandSize = n;
}
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;
}
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) {
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);
}