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Example 1 with PrecisionResultProcedure

use of uk.ac.sussex.gdsc.smlm.results.procedures.PrecisionResultProcedure in project GDSC-SMLM by aherbert.

the class CreateData method showSummary.

private double showSummary(List<? extends FluorophoreSequenceModel> fluorophores, List<LocalisationModel> localisations) {
    IJ.showStatus("Calculating statistics ...");
    final Statistics[] stats = new Statistics[NAMES.length];
    for (int i = 0; i < stats.length; i++) {
        stats[i] = (settings.getShowHistograms() || alwaysRemoveOutliers[i]) ? new StoredDataStatistics() : new Statistics();
    }
    // Find the largest timepoint
    final ImagePlus outputImp = WindowManager.getImage(benchmarkImageId);
    int frameCount;
    if (outputImp == null) {
        sortLocalisationsByTime(localisations);
        frameCount = localisations.get(localisations.size() - 1).getTime();
    } else {
        frameCount = outputImp.getStackSize();
    }
    final int[] countHistogram = new int[frameCount + 1];
    // Use the localisations that were drawn to create the sampled on/off times
    rebuildNeighbours(localisations);
    // Assume that there is at least one localisation
    final LocalisationModel first = localisations.get(0);
    // The current localisation
    int currentId = first.getId();
    // The last time this localisation was on
    int lastT = first.getTime();
    // Number of blinks
    int blinks = 0;
    // On-time of current pulse
    int currentT = 0;
    double signal = 0;
    final double centreOffset = settings.getSize() * 0.5;
    // Used to convert the sampled times in frames into seconds
    final double framesPerSecond = 1000.0 / settings.getExposureTime();
    // final double gain = new CreateDataSettingsHelper(settings).getTotalGainSafe();
    for (final LocalisationModel l : localisations) {
        final double[] data = l.getData();
        if (data == null) {
            throw new IllegalStateException("No localisation data. This should not happen!");
        }
        final double noise = data[1];
        final double sx = data[2];
        final double sy = data[3];
        final double intensityInPhotons = data[4];
        // Q. What if the noise is zero, i.e. no background photon / read noise?
        // Just ignore it at current. This is only an approximation to the SNR estimate
        // if this is not a Gaussian spot.
        final double snr = Gaussian2DPeakResultHelper.getMeanSignalUsingP05(intensityInPhotons, sx, sy) / noise;
        stats[SIGNAL].add(intensityInPhotons);
        stats[NOISE].add(noise);
        if (noise != 0) {
            stats[SNR].add(snr);
        }
        // if (l.isContinuous())
        if (l.getNext() != null && l.getPrevious() != null) {
            stats[SIGNAL_CONTINUOUS].add(intensityInPhotons);
            if (noise != 0) {
                stats[SNR_CONTINUOUS].add(snr);
            }
        }
        final int id = l.getId();
        // Check if this a new fluorophore
        if (currentId != id) {
            // Add previous fluorophore
            stats[SAMPLED_BLINKS].add(blinks);
            stats[SAMPLED_T_ON].add(currentT / framesPerSecond);
            stats[TOTAL_SIGNAL].add(signal);
            // Reset
            blinks = 0;
            currentT = 1;
            currentId = id;
            signal = intensityInPhotons;
        } else {
            signal += intensityInPhotons;
            // Check if the current fluorophore pulse is broken (i.e. a blink)
            if (l.getTime() - 1 > lastT) {
                blinks++;
                stats[SAMPLED_T_ON].add(currentT / framesPerSecond);
                currentT = 1;
                stats[SAMPLED_T_OFF].add(((l.getTime() - 1) - lastT) / framesPerSecond);
            } else {
                // Continuous on-time
                currentT++;
            }
        }
        lastT = l.getTime();
        countHistogram[lastT]++;
        stats[X].add((l.getX() - centreOffset) * settings.getPixelPitch());
        stats[Y].add((l.getY() - centreOffset) * settings.getPixelPitch());
        stats[Z].add(l.getZ() * settings.getPixelPitch());
    }
    // Final fluorophore
    stats[SAMPLED_BLINKS].add(blinks);
    stats[SAMPLED_T_ON].add(currentT / framesPerSecond);
    stats[TOTAL_SIGNAL].add(signal);
    // Samples per frame
    for (int t = 1; t < countHistogram.length; t++) {
        stats[SAMPLES].add(countHistogram[t]);
    }
    if (fluorophores != null) {
        for (final FluorophoreSequenceModel f : fluorophores) {
            stats[BLINKS].add(f.getNumberOfBlinks());
            // On-time
            for (final double t : f.getOnTimes()) {
                stats[T_ON].add(t);
            }
            // Off-time
            for (final double t : f.getOffTimes()) {
                stats[T_OFF].add(t);
            }
        }
    } else {
        // show no blinks
        stats[BLINKS].add(0);
        stats[T_ON].add(1);
    }
    if (results != null) {
        // Convert depth-of-field to pixels
        final double depth = settings.getDepthOfField() / settings.getPixelPitch();
        try {
            // Get widths
            final WidthResultProcedure wp = new WidthResultProcedure(results, DistanceUnit.PIXEL);
            wp.getW();
            stats[WIDTH].add(wp.wx);
        } catch (final DataException ex) {
            ImageJUtils.log("Unable to compute width: " + ex.getMessage());
        }
        try {
            // Get z depth
            final StandardResultProcedure sp = new StandardResultProcedure(results, DistanceUnit.PIXEL);
            sp.getXyz();
            // Get precision
            final PrecisionResultProcedure pp = new PrecisionResultProcedure(results);
            pp.getPrecision();
            stats[PRECISION].add(pp.precisions);
            for (int i = 0; i < pp.size(); i++) {
                if (Math.abs(sp.z[i]) < depth) {
                    stats[PRECISION_IN_FOCUS].add(pp.precisions[i]);
                }
            }
        } catch (final DataException ex) {
            ImageJUtils.log("Unable to compute LSE precision: " + ex.getMessage());
        }
        // Compute density per frame. Multi-thread for speed
        if (settings.getDensityRadius() > 0) {
            final int threadCount = Prefs.getThreads();
            final Ticker ticker = ImageJUtils.createTicker(results.getLastFrame(), threadCount, "Calculating density ...");
            final ExecutorService threadPool = Executors.newFixedThreadPool(threadCount);
            final List<Future<?>> futures = new LinkedList<>();
            final TFloatArrayList coordsX = new TFloatArrayList();
            final TFloatArrayList coordsY = new TFloatArrayList();
            final Statistics densityStats = stats[DENSITY];
            final float radius = (float) (settings.getDensityRadius() * getHwhm());
            final Rectangle bounds = results.getBounds();
            final double area = (double) bounds.width * bounds.height;
            // Store the density for each result.
            final int[] allDensity = new int[results.size()];
            final FrameCounter counter = results.newFrameCounter();
            results.forEach((PeakResultProcedure) result -> {
                if (counter.advance(result.getFrame())) {
                    counter.increment(runDensityCalculation(threadPool, futures, coordsX, coordsY, densityStats, radius, area, allDensity, counter.getCount(), ticker));
                }
                coordsX.add(result.getXPosition());
                coordsY.add(result.getYPosition());
            });
            runDensityCalculation(threadPool, futures, coordsX, coordsY, densityStats, radius, area, allDensity, counter.getCount(), ticker);
            ConcurrencyUtils.waitForCompletionUnchecked(futures);
            threadPool.shutdown();
            ImageJUtils.finished();
            // Split results into singles (density = 0) and clustered (density > 0)
            final MemoryPeakResults singles = copyMemoryPeakResults("No Density");
            final MemoryPeakResults clustered = copyMemoryPeakResults("Density");
            counter.reset();
            results.forEach((PeakResultProcedure) result -> {
                final int density = allDensity[counter.getAndIncrement()];
                result.setOrigValue(density);
                if (density == 0) {
                    singles.add(result);
                } else {
                    clustered.add(result);
                }
            });
        }
    }
    final StringBuilder sb = new StringBuilder();
    sb.append(datasetNumber).append('\t');
    if (settings.getCameraType() == CameraType.SCMOS) {
        sb.append("sCMOS (").append(settings.getCameraModelName()).append(") ");
        final Rectangle bounds = cameraModel.getBounds();
        sb.append(" ").append(bounds.x).append(",").append(bounds.y);
        final int size = settings.getSize();
        sb.append(" ").append(size).append("x").append(size);
    } else if (CalibrationProtosHelper.isCcdCameraType(settings.getCameraType())) {
        sb.append(CalibrationProtosHelper.getName(settings.getCameraType()));
        final int size = settings.getSize();
        sb.append(" ").append(size).append("x").append(size);
        if (settings.getCameraType() == CameraType.EMCCD) {
            sb.append(" EM=").append(settings.getEmGain());
        }
        sb.append(" CG=").append(settings.getCameraGain());
        sb.append(" RN=").append(settings.getReadNoise());
        sb.append(" B=").append(settings.getBias());
    } else {
        throw new IllegalStateException();
    }
    sb.append(" QE=").append(settings.getQuantumEfficiency()).append('\t');
    sb.append(settings.getPsfModel());
    if (psfModelType == PSF_MODEL_IMAGE) {
        sb.append(" Image").append(settings.getPsfImageName());
    } else if (psfModelType == PSF_MODEL_ASTIGMATISM) {
        sb.append(" model=").append(settings.getAstigmatismModel());
    } else {
        sb.append(" DoF=").append(MathUtils.rounded(settings.getDepthOfFocus()));
        if (settings.getEnterWidth()) {
            sb.append(" SD=").append(MathUtils.rounded(settings.getPsfSd()));
        } else {
            sb.append(" λ=").append(MathUtils.rounded(settings.getWavelength()));
            sb.append(" NA=").append(MathUtils.rounded(settings.getNumericalAperture()));
        }
    }
    sb.append('\t');
    sb.append((fluorophores == null) ? localisations.size() : fluorophores.size()).append('\t');
    sb.append(stats[SAMPLED_BLINKS].getN() + (int) stats[SAMPLED_BLINKS].getSum()).append('\t');
    sb.append(localisations.size()).append('\t');
    sb.append(frameCount).append('\t');
    sb.append(MathUtils.rounded(areaInUm)).append('\t');
    sb.append(MathUtils.rounded(localisations.size() / (areaInUm * frameCount), 4)).append('\t');
    sb.append(MathUtils.rounded(getHwhm(), 4)).append('\t');
    double sd = getPsfSd();
    sb.append(MathUtils.rounded(sd, 4)).append('\t');
    sd *= settings.getPixelPitch();
    final double sa = PsfCalculator.squarePixelAdjustment(sd, settings.getPixelPitch()) / settings.getPixelPitch();
    sb.append(MathUtils.rounded(sa, 4)).append('\t');
    // Width not valid for the Image PSF.
    // Q. Is this true? We can approximate the FHWM for a spot-like image PSF.
    final int nStats = (psfModelType == PSF_MODEL_IMAGE) ? stats.length - 1 : stats.length;
    for (int i = 0; i < nStats; i++) {
        final double centre = (alwaysRemoveOutliers[i]) ? ((StoredDataStatistics) stats[i]).getStatistics().getPercentile(50) : stats[i].getMean();
        sb.append(MathUtils.rounded(centre, 4)).append('\t');
    }
    createSummaryTable().accept(sb.toString());
    // Show histograms
    if (settings.getShowHistograms() && !java.awt.GraphicsEnvironment.isHeadless()) {
        IJ.showStatus("Calculating histograms ...");
        final boolean[] chosenHistograms = getChoosenHistograms();
        final WindowOrganiser wo = new WindowOrganiser();
        final HistogramPlotBuilder builder = new HistogramPlotBuilder(TITLE);
        for (int i = 0; i < NAMES.length; i++) {
            if (chosenHistograms[i]) {
                builder.setData((StoredDataStatistics) stats[i]).setName(NAMES[i]).setIntegerBins(integerDisplay[i]).setRemoveOutliersOption((settings.getRemoveOutliers() || alwaysRemoveOutliers[i]) ? 2 : 0).setNumberOfBins(settings.getHistogramBins()).show(wo);
            }
        }
        wo.tile();
    }
    IJ.showStatus("");
    return stats[SIGNAL].getMean();
}
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HistogramPlotBuilder(uk.ac.sussex.gdsc.core.ij.HistogramPlot.HistogramPlotBuilder) MemoryPeakResults(uk.ac.sussex.gdsc.smlm.results.MemoryPeakResults) ImmutableMemoryPeakResults(uk.ac.sussex.gdsc.smlm.results.ImmutableMemoryPeakResults) Ticker(uk.ac.sussex.gdsc.core.logging.Ticker) FrameCounter(uk.ac.sussex.gdsc.smlm.results.count.FrameCounter) StoredDataStatistics(uk.ac.sussex.gdsc.core.utils.StoredDataStatistics) WidthResultProcedure(uk.ac.sussex.gdsc.smlm.results.procedures.WidthResultProcedure) WindowOrganiser(uk.ac.sussex.gdsc.core.ij.plugin.WindowOrganiser) PrecisionResultProcedure(uk.ac.sussex.gdsc.smlm.results.procedures.PrecisionResultProcedure) SummaryStatistics(org.apache.commons.math3.stat.descriptive.SummaryStatistics) StoredDataStatistics(uk.ac.sussex.gdsc.core.utils.StoredDataStatistics) Statistics(uk.ac.sussex.gdsc.core.utils.Statistics) ImagePlus(ij.ImagePlus) ReadHint(uk.ac.sussex.gdsc.smlm.results.ImageSource.ReadHint) LinkedList(java.util.LinkedList) TFloatArrayList(gnu.trove.list.array.TFloatArrayList) DataException(uk.ac.sussex.gdsc.core.data.DataException) LocalisationModel(uk.ac.sussex.gdsc.smlm.model.LocalisationModel) FluorophoreSequenceModel(uk.ac.sussex.gdsc.smlm.model.FluorophoreSequenceModel) ExecutorService(java.util.concurrent.ExecutorService) Future(java.util.concurrent.Future) StandardResultProcedure(uk.ac.sussex.gdsc.smlm.results.procedures.StandardResultProcedure)

Example 2 with PrecisionResultProcedure

use of uk.ac.sussex.gdsc.smlm.results.procedures.PrecisionResultProcedure in project GDSC-SMLM by aherbert.

the class PcPalmMolecules method extractLocalisations.

/**
 * Extract molecules for the PC-PALM analysis.
 *
 * <p>Estimate the localisation uncertainty (precision) of each molecule using the formula of
 * Mortensen, et al (2010), Nature Methods 7, 377-381. Store distance in nm and signal in photons
 * using the calibration
 *
 * @param results the results
 * @return the array list
 * @throws DataException If conversion to nm and photons with computed precision is not possible
 */
public List<Molecule> extractLocalisations(MemoryPeakResults results) {
    final ArrayList<Molecule> list = new ArrayList<>(results.size());
    // Access calibrated data
    final StandardResultProcedure sp = new StandardResultProcedure(results, DistanceUnit.NM, IntensityUnit.PHOTON);
    sp.getIxy();
    final PrecisionResultProcedure pp = new PrecisionResultProcedure(results);
    pp.getPrecision();
    for (int i = 0, size = pp.size(); i < size; i++) {
        list.add(new Molecule(sp.x[i], sp.y[i], pp.precisions[i], sp.intensity[i]));
    }
    return list;
}
Also used : TDoubleArrayList(gnu.trove.list.array.TDoubleArrayList) ArrayList(java.util.ArrayList) StandardResultProcedure(uk.ac.sussex.gdsc.smlm.results.procedures.StandardResultProcedure) PrecisionResultProcedure(uk.ac.sussex.gdsc.smlm.results.procedures.PrecisionResultProcedure) WeightedObservedPoint(org.apache.commons.math3.fitting.WeightedObservedPoint) ClusterPoint(uk.ac.sussex.gdsc.core.clustering.ClusterPoint)

Example 3 with PrecisionResultProcedure

use of uk.ac.sussex.gdsc.smlm.results.procedures.PrecisionResultProcedure in project GDSC-SMLM by aherbert.

the class PcPalmMolecules method runManualTracing.

private void runManualTracing() {
    if (!showManualTracingDialog()) {
        return;
    }
    startLog();
    // Convert seconds to frames
    final int timeInFrames = Math.max(1, (int) Math.round(settings.timeThreshold * 1000.0 / settings.results.getCalibrationReader().getExposureTime()));
    // Get precisions
    final PrecisionResultProcedure pp = new PrecisionResultProcedure(settings.results);
    pp.getPrecision();
    final ArrayList<Molecule> singles = new ArrayList<>();
    settings.molecules = traceMolecules(settings.results, pp.precisions, settings.distanceThreshold, timeInFrames, singles);
    settings.molecules.addAll(singles);
}
Also used : TDoubleArrayList(gnu.trove.list.array.TDoubleArrayList) ArrayList(java.util.ArrayList) PrecisionResultProcedure(uk.ac.sussex.gdsc.smlm.results.procedures.PrecisionResultProcedure) WeightedObservedPoint(org.apache.commons.math3.fitting.WeightedObservedPoint) ClusterPoint(uk.ac.sussex.gdsc.core.clustering.ClusterPoint)

Example 4 with PrecisionResultProcedure

use of uk.ac.sussex.gdsc.smlm.results.procedures.PrecisionResultProcedure in project GDSC-SMLM by aherbert.

the class Fire method canCalculatePrecision.

private boolean canCalculatePrecision(MemoryPeakResults results) {
    try {
        pp = new PrecisionResultProcedure(results);
        pp.getLsePrecision();
    } catch (final DataException ex) {
        return false;
    }
    // Check they are different
    for (int i = 0; i < pp.size(); i++) {
        // Check this is valid
        if (Double.isFinite(pp.precisions[i])) {
            final double p1 = pp.precisions[i];
            for (int j = i + 1; j < pp.size(); j++) {
                if (Double.isFinite(pp.precisions[j]) && pp.precisions[j] != p1) {
                    return true;
                }
            }
            // All the results are the same, this is not valid
            break;
        }
    }
    return false;
}
Also used : DataException(uk.ac.sussex.gdsc.core.data.DataException) PrecisionResultProcedure(uk.ac.sussex.gdsc.smlm.results.procedures.PrecisionResultProcedure) WeightedObservedPoint(org.apache.commons.math3.fitting.WeightedObservedPoint)

Example 5 with PrecisionResultProcedure

use of uk.ac.sussex.gdsc.smlm.results.procedures.PrecisionResultProcedure in project GDSC-SMLM by aherbert.

the class ImageJ3DResultsViewer method createSphereSizeFromPrecision.

@Nullable
private static Point3f[] createSphereSizeFromPrecision(MemoryPeakResults results) {
    final PrecisionResultProcedure p = new PrecisionResultProcedure(results);
    try {
        final PrecisionMethod m = p.getPrecision();
        IJ.log("Using precision method " + FitProtosHelper.getName(m));
        final Point3f[] size = new Point3f[results.size()];
        for (int i = 0, j = 0; i < p.precisions.length; i++) {
            // Precision is in NM which matches the rendering
            final float v = (float) p.precisions[i];
            size[j++] = new Point3f(v, v, v);
        }
        return size;
    } catch (final DataException ex) {
        IJ.error(TITLE, "The results have no precision: " + ex.getMessage());
        return null;
    }
}
Also used : DataException(uk.ac.sussex.gdsc.core.data.DataException) Point3f(org.scijava.vecmath.Point3f) PrecisionResultProcedure(uk.ac.sussex.gdsc.smlm.results.procedures.PrecisionResultProcedure) PrecisionMethod(uk.ac.sussex.gdsc.smlm.data.config.FitProtos.PrecisionMethod) Nullable(uk.ac.sussex.gdsc.core.annotation.Nullable)

Aggregations

PrecisionResultProcedure (uk.ac.sussex.gdsc.smlm.results.procedures.PrecisionResultProcedure)11 DataException (uk.ac.sussex.gdsc.core.data.DataException)6 ArrayList (java.util.ArrayList)5 PeakResult (uk.ac.sussex.gdsc.smlm.results.PeakResult)5 Statistics (uk.ac.sussex.gdsc.core.utils.Statistics)4 StandardResultProcedure (uk.ac.sussex.gdsc.smlm.results.procedures.StandardResultProcedure)4 Rectangle (java.awt.Rectangle)3 WeightedObservedPoint (org.apache.commons.math3.fitting.WeightedObservedPoint)3 ClusterPoint (uk.ac.sussex.gdsc.core.clustering.ClusterPoint)3 StoredDataStatistics (uk.ac.sussex.gdsc.core.utils.StoredDataStatistics)3 TDoubleArrayList (gnu.trove.list.array.TDoubleArrayList)2 IJ (ij.IJ)2 ImagePlus (ij.ImagePlus)2 ImageStack (ij.ImageStack)2 WindowManager (ij.WindowManager)2 GenericDialog (ij.gui.GenericDialog)2 PlugIn (ij.plugin.PlugIn)2 FloatProcessor (ij.process.FloatProcessor)2 ImageProcessor (ij.process.ImageProcessor)2 DescriptiveStatistics (org.apache.commons.math3.stat.descriptive.DescriptiveStatistics)2