Examples with FluorophoreSequenceModel gdsc.smlm.model.FluorophoreSequenceModel used on opensource projects

Example 1 with FluorophoreSequenceModel

use of gdsc.smlm.model.FluorophoreSequenceModel in project GDSC-SMLM by aherbert.

the class CreateData method run.

/*
	 * (non-Javadoc)
	 * 
	 * @see ij.plugin.PlugIn#run(java.lang.String)
	 */
public void run(String arg) {
    SMLMUsageTracker.recordPlugin(this.getClass(), arg);
    extraOptions = Utils.isExtraOptions();
    simpleMode = (arg != null && arg.contains("simple"));
    benchmarkMode = (arg != null && arg.contains("benchmark"));
    spotMode = (arg != null && arg.contains("spot"));
    trackMode = (arg != null && arg.contains("track"));
    if ("load".equals(arg)) {
        loadBenchmarkData();
        return;
    }
    // Each localisation is a simulated emission of light from a point in space and time
    List<LocalisationModel> localisations = null;
    // Each localisation set is a collection of localisations that represent all localisations
    // with the same ID that are on in the same image time frame (Note: the simulation
    // can create many localisations per fluorophore per time frame which is useful when 
    // modelling moving particles)
    List<LocalisationModelSet> localisationSets = null;
    // Each fluorophore contains the on and off times when light was emitted 
    List<? extends FluorophoreSequenceModel> fluorophores = null;
    if (simpleMode || benchmarkMode || spotMode) {
        if (!showSimpleDialog())
            return;
        resetMemory();
        // 1 second frames
        settings.exposureTime = 1000;
        areaInUm = settings.size * settings.pixelPitch * settings.size * settings.pixelPitch / 1e6;
        // Number of spots per frame
        int n = 0;
        int[] nextN = null;
        SpatialDistribution dist;
        if (benchmarkMode) {
            // --------------------
            // BENCHMARK SIMULATION
            // --------------------
            // Draw the same point on the image repeatedly
            n = 1;
            dist = createFixedDistribution();
            reportAndSaveFittingLimits(dist);
        } else if (spotMode) {
            // ---------------
            // SPOT SIMULATION
            // ---------------
            // The spot simulation draws 0 or 1 random point per frame. 
            // Ensure we have 50% of the frames with a spot.
            nextN = new int[settings.particles * 2];
            Arrays.fill(nextN, 0, settings.particles, 1);
            Random rand = new Random();
            rand.shuffle(nextN);
            // Only put spots in the central part of the image
            double border = settings.size / 4.0;
            dist = createUniformDistribution(border);
        } else {
            // -----------------
            // SIMPLE SIMULATION
            // -----------------
            // The simple simulation draws n random points per frame to achieve a specified density.
            // No points will appear in multiple frames.
            // Each point has a random number of photons sampled from a range.
            // We can optionally use a mask. Create his first as it updates the areaInUm
            dist = createDistribution();
            // Randomly sample (i.e. not uniform density in all frames)
            if (settings.samplePerFrame) {
                final double mean = areaInUm * settings.density;
                System.out.printf("Mean samples = %f\n", mean);
                if (mean < 0.5) {
                    GenericDialog gd = new GenericDialog(TITLE);
                    gd.addMessage("The mean samples per frame is low: " + Utils.rounded(mean) + "\n \nContinue?");
                    gd.enableYesNoCancel();
                    gd.hideCancelButton();
                    gd.showDialog();
                    if (!gd.wasOKed())
                        return;
                }
                PoissonDistribution poisson = new PoissonDistribution(createRandomGenerator(), mean, PoissonDistribution.DEFAULT_EPSILON, PoissonDistribution.DEFAULT_MAX_ITERATIONS);
                StoredDataStatistics samples = new StoredDataStatistics(settings.particles);
                while (samples.getSum() < settings.particles) {
                    samples.add(poisson.sample());
                }
                nextN = new int[samples.getN()];
                for (int i = 0; i < nextN.length; i++) nextN[i] = (int) samples.getValue(i);
            } else {
                // Use the density to get the number per frame
                n = (int) FastMath.max(1, Math.round(areaInUm * settings.density));
            }
        }
        RandomGenerator random = null;
        localisations = new ArrayList<LocalisationModel>(settings.particles);
        localisationSets = new ArrayList<LocalisationModelSet>(settings.particles);
        final int minPhotons = (int) settings.photonsPerSecond;
        final int range = (int) settings.photonsPerSecondMaximum - minPhotons + 1;
        if (range > 1)
            random = createRandomGenerator();
        // Add frames at the specified density until the number of particles has been reached
        int id = 0;
        int t = 0;
        while (id < settings.particles) {
            // Allow the number per frame to be specified
            if (nextN != null) {
                if (t >= nextN.length)
                    break;
                n = nextN[t];
            }
            // Simulate random positions in the frame for the specified density
            t++;
            for (int j = 0; j < n; j++) {
                final double[] xyz = dist.next();
                // Ignore within border. We do not want to draw things we cannot fit.
                //if (!distBorder.isWithinXY(xyz))
                //	continue;
                // Simulate random photons
                final int intensity = minPhotons + ((random != null) ? random.nextInt(range) : 0);
                LocalisationModel m = new LocalisationModel(id, t, xyz, intensity, LocalisationModel.CONTINUOUS);
                localisations.add(m);
                // Each localisation can be a separate localisation set
                LocalisationModelSet set = new LocalisationModelSet(id, t);
                set.add(m);
                localisationSets.add(set);
                id++;
            }
        }
    } else {
        if (!showDialog())
            return;
        resetMemory();
        areaInUm = settings.size * settings.pixelPitch * settings.size * settings.pixelPitch / 1e6;
        int totalSteps;
        double correlation = 0;
        ImageModel imageModel;
        if (trackMode) {
            // ----------------
            // TRACK SIMULATION
            // ----------------
            // In track mode we create fixed lifetime fluorophores that do not overlap in time.
            // This is the simplest simulation to test moving molecules.
            settings.seconds = (int) Math.ceil(settings.particles * (settings.exposureTime + settings.tOn) / 1000);
            totalSteps = 0;
            final double simulationStepsPerFrame = (settings.stepsPerSecond * settings.exposureTime) / 1000.0;
            imageModel = new FixedLifetimeImageModel(settings.stepsPerSecond * settings.tOn / 1000.0, simulationStepsPerFrame);
        } else {
            // ---------------
            // FULL SIMULATION
            // ---------------
            // The full simulation draws n random points in space.
            // The same molecule may appear in multiple frames, move and blink.
            //
            // Points are modelled as fluorophores that must be activated and then will 
            // blink and photo-bleach. The molecules may diffuse and this can be simulated 
            // with many steps per image frame. All steps from a frame are collected
            // into a localisation set which can be drawn on the output image.
            SpatialIllumination activationIllumination = createIllumination(settings.pulseRatio, settings.pulseInterval);
            // Generate additional frames so that each frame has the set number of simulation steps
            totalSteps = (int) Math.ceil(settings.seconds * settings.stepsPerSecond);
            // Since we have an exponential decay of activations
            // ensure half of the particles have activated by 30% of the frames.
            double eAct = totalSteps * 0.3 * activationIllumination.getAveragePhotons();
            // Q. Does tOn/tOff change depending on the illumination strength?
            imageModel = new ActivationEnergyImageModel(eAct, activationIllumination, settings.stepsPerSecond * settings.tOn / 1000.0, settings.stepsPerSecond * settings.tOffShort / 1000.0, settings.stepsPerSecond * settings.tOffLong / 1000.0, settings.nBlinksShort, settings.nBlinksLong);
            imageModel.setUseGeometricDistribution(settings.nBlinksGeometricDistribution);
            // Only use the correlation if selected for the distribution
            if (PHOTON_DISTRIBUTION[PHOTON_CORRELATED].equals(settings.photonDistribution))
                correlation = settings.correlation;
        }
        imageModel.setRandomGenerator(createRandomGenerator());
        imageModel.setPhotonBudgetPerFrame(true);
        imageModel.setDiffusion2D(settings.diffuse2D);
        imageModel.setRotation2D(settings.rotate2D);
        IJ.showStatus("Creating molecules ...");
        SpatialDistribution distribution = createDistribution();
        List<CompoundMoleculeModel> compounds = createCompoundMolecules();
        if (compounds == null)
            return;
        List<CompoundMoleculeModel> molecules = imageModel.createMolecules(compounds, settings.particles, distribution, settings.rotateInitialOrientation);
        // Activate fluorophores
        IJ.showStatus("Creating fluorophores ...");
        // Note: molecules list will be converted to compounds containing fluorophores
        fluorophores = imageModel.createFluorophores(molecules, totalSteps);
        if (fluorophores.isEmpty()) {
            IJ.error(TITLE, "No fluorophores created");
            return;
        }
        IJ.showStatus("Creating localisations ...");
        // TODO - Output a molecule Id for each fluorophore if using compound molecules. This allows analysis
        // of the ratio of trimers, dimers, monomers, etc that could be detected.
        totalSteps = checkTotalSteps(totalSteps, fluorophores);
        if (totalSteps == 0)
            return;
        imageModel.setPhotonDistribution(createPhotonDistribution());
        imageModel.setConfinementDistribution(createConfinementDistribution());
        // This should be optimised
        imageModel.setConfinementAttempts(10);
        localisations = imageModel.createImage(molecules, settings.fixedFraction, totalSteps, (double) settings.photonsPerSecond / settings.stepsPerSecond, correlation, settings.rotateDuringSimulation);
        // Re-adjust the fluorophores to the correct time
        if (settings.stepsPerSecond != 1) {
            final double scale = 1.0 / settings.stepsPerSecond;
            for (FluorophoreSequenceModel f : fluorophores) f.adjustTime(scale);
        }
        // Integrate the frames
        localisationSets = combineSimulationSteps(localisations);
        localisationSets = filterToImageBounds(localisationSets);
    }
    datasetNumber++;
    localisations = drawImage(localisationSets);
    if (localisations == null || localisations.isEmpty()) {
        IJ.error(TITLE, "No localisations created");
        return;
    }
    fluorophores = removeFilteredFluorophores(fluorophores, localisations);
    double signalPerFrame = showSummary(fluorophores, localisations);
    if (!benchmarkMode) {
        boolean fullSimulation = (!(simpleMode || spotMode));
        saveSimulationParameters(localisations.size(), fullSimulation, signalPerFrame);
    }
    IJ.showStatus("Saving data ...");
    //convertRelativeToAbsolute(molecules);
    saveFluorophores(fluorophores);
    saveImageResults(results);
    saveLocalisations(localisations);
    // The settings for the filenames may have changed
    SettingsManager.saveSettings(globalSettings);
    IJ.showStatus("Done");
}

Example 2 with FluorophoreSequenceModel

use of gdsc.smlm.model.FluorophoreSequenceModel in project GDSC-SMLM by aherbert.

the class CreateData method removeFilteredFluorophores.

/**
	 * Remove all fluorophores which were not drawn
	 * 
	 * @param fluorophores
	 * @param localisations
	 * @return
	 */
private List<? extends FluorophoreSequenceModel> removeFilteredFluorophores(List<? extends FluorophoreSequenceModel> fluorophores, List<LocalisationModel> localisations) {
    if (fluorophores == null)
        return null;
    // movingMolecules will be created with an initial capacity to hold all the unique IDs
    TIntHashSet idSet = new TIntHashSet((movingMolecules != null) ? movingMolecules.capacity() : 0);
    for (LocalisationModel l : localisations) idSet.add(l.getId());
    List<FluorophoreSequenceModel> newFluorophores = new ArrayList<FluorophoreSequenceModel>(idSet.size());
    for (FluorophoreSequenceModel f : fluorophores) {
        if (idSet.contains(f.getId()))
            newFluorophores.add(f);
    }
    return newFluorophores;
}

Example 3 with FluorophoreSequenceModel

use of gdsc.smlm.model.FluorophoreSequenceModel 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 ...");
    createSummaryTable();
    Statistics[] stats = new Statistics[NAMES.length];
    for (int i = 0; i < stats.length; i++) {
        stats[i] = (settings.showHistograms || alwaysRemoveOutliers[i]) ? new StoredDataStatistics() : new Statistics();
    }
    // Find the largest timepoint
    ImagePlus outputImp = WindowManager.getImage(benchmarkImageId);
    int nFrames;
    if (outputImp == null) {
        sortLocalisationsByTime(localisations);
        nFrames = localisations.get(localisations.size() - 1).getTime();
    } else {
        nFrames = outputImp.getStackSize();
    }
    int[] countHistogram = new int[nFrames + 1];
    // Use the localisations that were drawn to create the sampled on/off times
    rebuildNeighbours(localisations);
    // Assume that there is at least one localisation
    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.size * 0.5;
    // Used to convert the sampled times in frames into seconds
    final double framesPerSecond = 1000.0 / settings.exposureTime;
    final double gain = (settings.getTotalGain() > 0) ? settings.getTotalGain() : 1;
    for (LocalisationModel l : localisations) {
        if (l.getData() == null)
            System.out.println("No localisation data. This should not happen!");
        final double noise = (l.getData() != null) ? l.getData()[1] : 1;
        final double intensity = (l.getData() != null) ? l.getData()[4] : l.getIntensity();
        final double intensityInPhotons = intensity / gain;
        // Q. What if the noise is zero, i.e. no background photon / read noise?
        // Just ignore it at current.
        final double snr = intensity / noise;
        stats[SIGNAL].add(intensityInPhotons);
        stats[NOISE].add(noise / gain);
        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);
        }
        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.pixelPitch);
        stats[Y].add((l.getY() - centreOffset) * settings.pixelPitch);
        stats[Z].add(l.getZ() * settings.pixelPitch);
    }
    // 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 (FluorophoreSequenceModel f : fluorophores) {
            stats[BLINKS].add(f.getNumberOfBlinks());
            // On-time
            for (double t : f.getOnTimes()) stats[T_ON].add(t);
            // Off-time
            for (double t : f.getOffTimes()) stats[T_OFF].add(t);
        }
    } else {
        // show no blinks
        stats[BLINKS].add(0);
        stats[T_ON].add(1);
    //stats[T_OFF].add(0);
    }
    if (results != null) {
        final boolean emCCD = (settings.getEmGain() > 1);
        // Convert depth-of-field to pixels
        final double depth = settings.depthOfField / settings.pixelPitch;
        for (PeakResult r : results.getResults()) {
            final double precision = r.getPrecision(settings.pixelPitch, gain, emCCD);
            stats[PRECISION].add(precision);
            // The error stores the z-depth in pixels
            if (Math.abs(r.error) < depth)
                stats[PRECISION_IN_FOCUS].add(precision);
            stats[WIDTH].add(r.getSD());
        }
        // Compute density per frame. Multithread for speed
        if (settings.densityRadius > 0) {
            IJ.showStatus("Calculating density ...");
            ExecutorService threadPool = Executors.newFixedThreadPool(Prefs.getThreads());
            List<Future<?>> futures = new LinkedList<Future<?>>();
            final ArrayList<float[]> coords = new ArrayList<float[]>();
            int t = results.getHead().getFrame();
            final Statistics densityStats = stats[DENSITY];
            final float radius = (float) (settings.densityRadius * getHWHM());
            final Rectangle bounds = results.getBounds();
            currentIndex = 0;
            finalIndex = results.getTail().getFrame();
            // Store the density for each result.
            int[] allDensity = new int[results.size()];
            int allIndex = 0;
            for (PeakResult r : results.getResults()) {
                if (t != r.getFrame()) {
                    allIndex += runDensityCalculation(threadPool, futures, coords, densityStats, radius, bounds, allDensity, allIndex);
                }
                coords.add(new float[] { r.getXPosition(), r.getYPosition() });
                t = r.getFrame();
            }
            runDensityCalculation(threadPool, futures, coords, densityStats, radius, bounds, allDensity, allIndex);
            Utils.waitForCompletion(futures);
            threadPool.shutdownNow();
            threadPool = null;
            IJ.showProgress(1);
            // Split results into singles (density = 0) and clustered (density > 0)
            MemoryPeakResults singles = copyMemoryPeakResults("No Density");
            MemoryPeakResults clustered = copyMemoryPeakResults("Density");
            int i = 0;
            for (PeakResult r : results.getResults()) {
                // Store density in the original value field
                r.origValue = allDensity[i];
                if (allDensity[i++] == 0)
                    singles.add(r);
                else
                    clustered.add(r);
            }
        }
    }
    StringBuilder sb = new StringBuilder();
    sb.append(datasetNumber).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(nFrames).append("\t");
    sb.append(Utils.rounded(areaInUm)).append("\t");
    sb.append(Utils.rounded(localisations.size() / (areaInUm * nFrames), 4)).append("\t");
    sb.append(Utils.rounded(getHWHM(), 4)).append("\t");
    double s = getPsfSD();
    sb.append(Utils.rounded(s, 4)).append("\t");
    s *= settings.pixelPitch;
    final double sa = PSFCalculator.squarePixelAdjustment(s, settings.pixelPitch) / settings.pixelPitch;
    sb.append(Utils.rounded(sa, 4)).append("\t");
    // Width not valid for the Image PSF
    int nStats = (imagePSF) ? stats.length - 1 : stats.length;
    for (int i = 0; i < nStats; i++) {
        double centre = (alwaysRemoveOutliers[i]) ? ((StoredDataStatistics) stats[i]).getStatistics().getPercentile(50) : stats[i].getMean();
        sb.append(Utils.rounded(centre, 4)).append("\t");
    }
    if (java.awt.GraphicsEnvironment.isHeadless()) {
        IJ.log(sb.toString());
        return stats[SIGNAL].getMean();
    } else {
        summaryTable.append(sb.toString());
    }
    // Show histograms
    if (settings.showHistograms) {
        IJ.showStatus("Calculating histograms ...");
        boolean[] chosenHistograms = getChoosenHistograms();
        WindowOrganiser wo = new WindowOrganiser();
        boolean requireRetile = false;
        for (int i = 0; i < NAMES.length; i++) {
            if (chosenHistograms[i]) {
                wo.add(Utils.showHistogram(TITLE, (StoredDataStatistics) stats[i], NAMES[i], (integerDisplay[i]) ? 1 : 0, (settings.removeOutliers || alwaysRemoveOutliers[i]) ? 2 : 0, settings.histogramBins * ((integerDisplay[i]) ? 100 : 1)));
                requireRetile = requireRetile || Utils.isNewWindow();
            }
        }
        wo.tile();
    }
    IJ.showStatus("");
    return stats[SIGNAL].getMean();
}

Example 4 with FluorophoreSequenceModel

use of gdsc.smlm.model.FluorophoreSequenceModel in project GDSC-SMLM by aherbert.

the class CreateData method checkTotalSteps.

/**
	 * Check if the total steps can fit all the fluorophores end times. If not then ask the user if they want to draw
	 * extra
	 * frames. Return the total steps to simulate (either the original steps or a larger number to fit all the data).
	 * 
	 * @param totalSteps
	 * @param fluorophores
	 * @return The new total steps to simulate
	 */
private int checkTotalSteps(int totalSteps, List<? extends FluorophoreSequenceModel> fluorophores) {
    int max = totalSteps;
    for (FluorophoreSequenceModel f : fluorophores) {
        if (max < f.getEndTime())
            max = (int) (f.getEndTime() + 1);
    }
    if (max > totalSteps) {
        GenericDialog gd = new GenericDialog(TITLE);
        gd.enableYesNoCancel();
        gd.hideCancelButton();
        final double simulationStepsPerFrame = (settings.stepsPerSecond * settings.exposureTime) / 1000.0;
        int newFrames = 1 + (int) (max / simulationStepsPerFrame);
        if (totalSteps != 0) {
            int totalFrames = (int) Math.ceil(settings.seconds * 1000 / settings.exposureTime);
            gd.addMessage(String.format("Require %d (%s%%) additional frames to draw all fluorophores.\nDo you want to add extra frames?", newFrames - totalFrames, Utils.rounded((100.0 * (newFrames - totalFrames)) / totalFrames, 3)));
        } else {
            gd.addMessage(String.format("Require %d frames to draw all fluorophores.\nDo you want to proceed?", newFrames));
        }
        gd.showDialog();
        if (gd.wasOKed())
            totalSteps = max;
    }
    return totalSteps;
}

Example 5 with FluorophoreSequenceModel

use of gdsc.smlm.model.FluorophoreSequenceModel in project GDSC-SMLM by aherbert.

the class CreateData method saveFluorophores.

/**
	 * Save the fluorophores to a text file
	 * 
	 * @param fluorophores
	 */
private void saveFluorophores(List<? extends FluorophoreSequenceModel> fluorophores) {
    if (!settings.saveFluorophores || fluorophores == null)
        return;
    String[] path = Utils.decodePath(settings.fluorophoresFilename);
    OpenDialog chooser = new OpenDialog("Fluorophores_File", path[0], path[1]);
    if (chooser.getFileName() != null) {
        settings.fluorophoresFilename = chooser.getDirectory() + chooser.getFileName();
        settings.fluorophoresFilename = Utils.replaceExtension(settings.fluorophoresFilename, "xls");
        BufferedWriter output = null;
        try {
            output = new BufferedWriter(new FileWriter(settings.fluorophoresFilename));
            output.write(createResultsFileHeader());
            output.write("#Id\tn-Blinks\tStart\tStop\t...");
            output.newLine();
            for (int id = 1; id <= fluorophores.size(); id++) {
                FluorophoreSequenceModel f = fluorophores.get(id - 1);
                StringBuffer sb = new StringBuffer();
                sb.append(f.getId()).append("\t");
                sb.append(f.getNumberOfBlinks()).append("\t");
                for (double[] burst : f.getBurstSequence()) {
                    sb.append(Utils.rounded(burst[0], 3)).append("\t").append(Utils.rounded(burst[1], 3)).append("\t");
                }
                output.write(sb.toString());
                output.newLine();
            }
        } catch (Exception e) {
            // Q. Add better handling of errors?
            e.printStackTrace();
            IJ.log("Failed to save fluorophores to file: " + settings.fluorophoresFilename);
        } finally {
            if (output != null) {
                try {
                    output.close();
                } catch (IOException e) {
                    e.printStackTrace();
                }
            }
        }
    }
}