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ModularImageAnalysis (MIA) is an ImageJ plugin which provides a modular framework for assembling image and object analysis workflows. Detected objects can be transformed, filtered, measured and related. Analysis workflows are batch-enabled by default, allowing easy processing of high-content datasets.

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// TODO: Show original and fit PSFs - maybe as a mosaic - to demonstrate the processAutomatic is working correctly

package io.github.mianalysis.mia.module.objects.process;


import java.util.Iterator;

import org.scijava.Priority;
import org.scijava.plugin.Plugin;

import com.drew.lang.annotations.Nullable;

import ij.IJ;
import ij.ImagePlus;
import ij.plugin.Duplicator;
import ij.process.ImageProcessor;
import io.github.mianalysis.mia.module.Categories;
import io.github.mianalysis.mia.module.Category;
import io.github.mianalysis.mia.module.Module;
import io.github.mianalysis.mia.module.Modules;
import io.github.mianalysis.mia.module.images.process.ImageMath;
import io.github.mianalysis.mia.module.images.transform.CropImage;
import io.github.mianalysis.mia.object.Measurement;
import io.github.mianalysis.mia.object.Obj;
import io.github.mianalysis.mia.object.Objs;
import io.github.mianalysis.mia.object.Workspace;
import io.github.mianalysis.mia.object.coordinates.Point;
import io.github.mianalysis.mia.object.image.Image;
import io.github.mianalysis.mia.object.image.ImageFactory;
import io.github.mianalysis.mia.object.parameters.BooleanP;
import io.github.mianalysis.mia.object.parameters.ChoiceP;
import io.github.mianalysis.mia.object.parameters.InputImageP;
import io.github.mianalysis.mia.object.parameters.InputObjectsP;
import io.github.mianalysis.mia.object.parameters.ObjectMeasurementP;
import io.github.mianalysis.mia.object.parameters.OutputImageP;
import io.github.mianalysis.mia.object.parameters.Parameters;
import io.github.mianalysis.mia.object.parameters.SeparatorP;
import io.github.mianalysis.mia.object.parameters.text.DoubleP;
import io.github.mianalysis.mia.object.parameters.text.IntegerP;
import io.github.mianalysis.mia.object.refs.ObjMeasurementRef;
import io.github.mianalysis.mia.object.refs.collections.ImageMeasurementRefs;
import io.github.mianalysis.mia.object.refs.collections.MetadataRefs;
import io.github.mianalysis.mia.object.refs.collections.ObjMeasurementRefs;
import io.github.mianalysis.mia.object.refs.collections.ObjMetadataRefs;
import io.github.mianalysis.mia.object.refs.collections.ParentChildRefs;
import io.github.mianalysis.mia.object.refs.collections.PartnerRefs;
import io.github.mianalysis.mia.object.system.Status;
import io.github.mianalysis.mia.process.imagej.GaussianFitter;
import io.github.mianalysis.mia.process.math.CumStat;
import io.github.mianalysis.mia.process.math.GaussianDistribution2D;

/**
 * Created by sc13967 on 05/06/2017.
 */

/**
* Gaussian spot fitting.  Can take objects as estimated locations.
***Only works in 2D***
***Only works for refinement of existing spots***
*/
@Plugin(type = Module.class, priority=Priority.LOW, visible=true)
public class FitGaussian2D extends Module {

	/**
	* 
	*/
    public static final String INPUT_SEPARATOR = "Image/objects input";

	/**
	* Image from the workspace to which the 2D Gaussian profiles will be fit.
	*/
    public static final String INPUT_IMAGE = "Input image";

	/**
	* Objects for which 2D Gaussians will be fit.  A single Gaussian will be calculated for each object, with the centroid of the input object used as the starting estimate for the Gaussian centroid.  Fit parameters will be stored as measurements associated with the relevant input objects.
	*/
    public static final String INPUT_OBJECTS = "Input objects";


	/**
	* 
	*/
    public static final String FITTING_SEPARATOR = "Fitting controls";

	/**
	* Controls how the initial estimate of Gaussian sigma will be calculated.  Note: The initial radius value can also influence the range of accepted sigma values (see "Limit sigma range" parameter).
  • "Fixed value" The same initial sigma value is applied to the fitting of all objects. This value is provided by the "Sigma" parameter.
  • "Measurement" A different sigma value is applied to the fitting of each object. This value is provided by the measurement (specified by "Sigma measurement") associated with the object being fit.
*/ public static final String SIGMA_MODE = "Method to estimate sigma"; /** * If "Method to estimate sigma" is set to "Fixed value", this is the initial sigma value that will be used in the fitting of all objects. */ public static final String SIGMA = "Sigma"; /** * If "Method to estimate sigma" is set to "Measurement", this is the measurement that will provide the intial sigma value used in the fitting of each object. */ public static final String SIGMA_MEASUREMENT = "Sigma measurement"; /** * If using object-associated measurements as the intial estimate of sigma, these measurements can be systematically increased or decreased with this multiplier. Values less than 1 will reduce the sigma estimate, while values greater than 1 will increase it. To use the default sigma value, set this to 1. */ public static final String MEASUREMENT_MULTIPLIER = "Measurement multiplier"; /** * When selected, the final sigma value must lie within a specific range. This range is controlled by the "Minimum sigma (multiplier)" and "Maximum sigma (multiplier)" parameters. */ public static final String LIMIT_SIGMA_RANGE = "Limit sigma range"; public static final String MIN_SIGMA = "Minimum sigma (multiplier)"; public static final String MAX_SIGMA = "Maximum sigma (multiplier)"; /** * When selected, the size of the cropped image region, to which the 2D Gaussian is fit, is controlled by the "Window size" parameter. Otherwise, the window size is automatically selected as 4 times the initial estimate of sigma, plus 1. */ public static final String FIXED_FITTING_WINDOW = "Fixed fitting window"; /** * If using a fixed fitting window ("Fixed fitting window" parameter selected), this is the size of the image crop (centered on the object being fit) that will be used. */ public static final String WINDOW_SIZE = "Window size"; /** * The maximum number of iterations the fitting algorithm can use. */ public static final String MAX_EVALUATIONS = "Maximum number of evaluations"; /** * */ public static final String OUTPUT_SEPARATOR = "Output controls"; /** * If selected, any objects for which fits can't be determined (fitter runs out of evaluations before completing) will be removed from the input object set. */ public static final String REMOVE_UNFIT = "Remove objects with failed fitting"; /** * If selected, the coordinates of the input objects will be updated to be circles. The output circles will be centred on the best-fit location and have radii equal to the x-axis sigma value. */ public static final String APPLY_VOLUME = "Apply volume"; /** * If selected, an image containing all calculated Gaussian fits will be added to the workspace. */ public static final String CREATE_GAUSSIAN_IMAGE = "Create Gaussian image"; /** * If "Create Gaussian image" is selected, this is the name of the output Gaussian profile image that will be added to the workspace. */ public static final String GAUSSIAN_IMAGE = "Gaussian image name"; public FitGaussian2D(Modules modules) { super("Fit Gaussian 2D", modules); } public interface SigmaModes { String FIXED_VALUE = "Fixed value"; String MEASUREMENT = "Measurement"; String[] ALL = new String[] { FIXED_VALUE, MEASUREMENT }; } public interface Measurements { String X0_PX = "GAUSSFIT2D // X0_(PX)"; String Y0_PX = "GAUSSFIT2D // Y0_(PX)"; String Z0_SLICE = "GAUSSFIT2D // Z0_(SLICE)_(CENTROID)"; String SIGMA_X_PX = "GAUSSFIT2D // SIGMA_X_(PX)"; String SIGMA_Y_PX = "GAUSSFIT2D // SIGMA_Y_(PX)"; String SIGMA_MEAN_PX = "GAUSSFIT2D // SIGMA_MEAN_(PX)"; String X0_CAL = "GAUSSFIT2D // X0_(${SCAL})"; String Y0_CAL = "GAUSSFIT2D // Y0_(${SCAL})"; String Z0_CAL = "GAUSSFIT2D // Z0_(${SCAL})_(CENTROID)"; String SIGMA_X_CAL = "GAUSSFIT2D // SIGMA_X_(${SCAL})"; String SIGMA_Y_CAL = "GAUSSFIT2D // SIGMA_Y_(${SCAL})"; String SIGMA_MEAN_CAL = "GAUSSFIT2D // SIGMA_MEAN_(${SCAL})"; String A_0 = "GAUSSFIT2D // A_0"; String A_BG = "GAUSSFIT2D // A_BG"; String THETA = "GAUSSFIT2D // THETA"; String ELLIPTICITY = "GAUSSFIT2D // ELLIPTICITY"; String RESIDUAL = "GAUSSFIT2D // RESIDUAL_(NORM)"; } static double[][] getLimits(int halfW, int r, boolean limitSigma, double minSigma, double maxSigma) { if (limitSigma) { minSigma = r * minSigma; maxSigma = r * maxSigma; } else { minSigma = 0; maxSigma = Double.MAX_VALUE; } return new double[][] { { 0, 2 * halfW + 1 }, { 0, 2 * halfW + 1 }, { minSigma, maxSigma }, // Sigma can't go to // zero { minSigma, maxSigma }, { -Double.MAX_VALUE, Double.MAX_VALUE }, { -Double.MAX_VALUE, Double.MAX_VALUE }, { 0, 2 * Math.PI } }; } static double[] estimateParameters(ImageProcessor iprCrop, int r) { double x0 = iprCrop.getWidth() / 2; // centroid x double y0 = iprCrop.getHeight() / 2; // centroid y double A0 = iprCrop.getStatistics().max; // peak amplitude double ABG = iprCrop.getStatistics().min; // background amplitude double th = Math.PI; // theta return new double[] { x0, y0, r, r, A0, ABG, th }; } static double calculateResidual(ImageProcessor iprCrop, double[] p) { GaussianDistribution2D fitDistribution2D = new GaussianDistribution2D(p[0], p[1], p[2], p[3], p[4], p[5], p[6]); GaussianDistribution2D offsetDistribution2D = new GaussianDistribution2D(p[0], p[1], p[2], p[3], p[4] - p[5], 0, p[6]); double residual = 0; double totalReal = 0; for (int xPos = 0; xPos < iprCrop.getWidth(); xPos++) { for (int yPos = 0; yPos < iprCrop.getHeight(); yPos++) { double realVal = iprCrop.get(xPos, yPos); double fitVal = fitDistribution2D.getValues(xPos, yPos)[0]; double offsetVal = offsetDistribution2D.getValues(xPos, yPos)[0]; residual = residual + Math.abs(realVal - fitVal); totalReal = totalReal + offsetVal; } } return residual / totalReal; } static void assignVolume(Objs objects) { // Replacing spot volumes with explicit volume Objs tempObjects = new Objs("SpotVolume", objects); for (Obj spotObject : objects.values()) { int radius = (int) Math.round(spotObject.getMeasurement(Measurements.SIGMA_X_PX).getValue()); Point cent = spotObject.getMeanCentroid(true, false); int[] centroid = new int[] { (int) Math.round(cent.getX()), (int) Math.round(cent.getY()), (int) Math.round(cent.getZ()) }; Obj volumeObject = GetLocalObjectRegion.getLocalRegion(spotObject, tempObjects, centroid, radius, false); spotObject.getCoordinateSet().clear(); spotObject.getCoordinateSet().addAll(volumeObject.getCoordinateSet()); spotObject.clearSurface(); spotObject.clearCentroid(); spotObject.clearProjected(); spotObject.clearROIs(); } } static void assignMissingMeasurements(Obj obj) { obj.addMeasurement(new Measurement(Measurements.X0_PX, Double.NaN)); obj.addMeasurement(new Measurement(Measurements.Y0_PX, Double.NaN)); obj.addMeasurement(new Measurement(Measurements.Z0_SLICE, Double.NaN)); obj.addMeasurement(new Measurement(Measurements.SIGMA_X_PX, Double.NaN)); obj.addMeasurement(new Measurement(Measurements.SIGMA_Y_PX, Double.NaN)); obj.addMeasurement(new Measurement(Measurements.SIGMA_MEAN_PX, Double.NaN)); obj.addMeasurement(new Measurement(Measurements.X0_CAL, Double.NaN)); obj.addMeasurement(new Measurement(Measurements.Y0_CAL, Double.NaN)); obj.addMeasurement(new Measurement(Measurements.Z0_CAL, Double.NaN)); obj.addMeasurement(new Measurement(Measurements.SIGMA_X_CAL, Double.NaN)); obj.addMeasurement(new Measurement(Measurements.SIGMA_Y_CAL, Double.NaN)); obj.addMeasurement(new Measurement(Measurements.SIGMA_MEAN_CAL, Double.NaN)); obj.addMeasurement(new Measurement(Measurements.A_0, Double.NaN)); obj.addMeasurement(new Measurement(Measurements.A_BG, Double.NaN)); obj.addMeasurement(new Measurement(Measurements.THETA, Double.NaN)); obj.addMeasurement(new Measurement(Measurements.ELLIPTICITY, Double.NaN)); obj.addMeasurement(new Measurement(Measurements.RESIDUAL, Double.NaN)); } static void assignMeasurements(Obj obj, ImageProcessor iprCrop, double[] p, int x, int y, int halfW) { double distPerPxXY = obj.getDppXY(); double distPerPxZ = obj.getDppZ(); double x0 = p[0] + x - halfW; double y0 = p[1] + y - halfW; double z0 = obj.getZMean(true, false); double sx = p[2]; double sy = p[3]; double A0 = p[4]; double ABG = p[5]; double th = p[6]; double residual = calculateResidual(iprCrop, p); double ellipticity = sx > sy ? (sx - sy) / sx : (sy - sx) / sy; double sm = (sx + sy) / 2; // Storing the results as measurements obj.addMeasurement(new Measurement(Measurements.X0_PX, x0)); obj.addMeasurement(new Measurement(Measurements.Y0_PX, y0)); obj.addMeasurement(new Measurement(Measurements.Z0_SLICE, z0)); obj.addMeasurement(new Measurement(Measurements.SIGMA_X_PX, sx)); obj.addMeasurement(new Measurement(Measurements.SIGMA_Y_PX, sy)); obj.addMeasurement(new Measurement(Measurements.SIGMA_MEAN_PX, sm)); obj.addMeasurement(new Measurement(Measurements.X0_CAL, x0 * distPerPxXY)); obj.addMeasurement(new Measurement(Measurements.Y0_CAL, y0 * distPerPxXY)); obj.addMeasurement(new Measurement(Measurements.Z0_CAL, z0 * distPerPxZ)); obj.addMeasurement(new Measurement(Measurements.SIGMA_X_CAL, sx * distPerPxXY)); obj.addMeasurement(new Measurement(Measurements.SIGMA_Y_CAL, sy * distPerPxXY)); obj.addMeasurement(new Measurement(Measurements.SIGMA_MEAN_CAL, sm * distPerPxXY)); obj.addMeasurement(new Measurement(Measurements.A_0, A0)); obj.addMeasurement(new Measurement(Measurements.A_BG, ABG)); obj.addMeasurement(new Measurement(Measurements.THETA, th)); obj.addMeasurement(new Measurement(Measurements.ELLIPTICITY, ellipticity)); obj.addMeasurement(new Measurement(Measurements.RESIDUAL, residual)); } static Image createGaussianImage(Objs objects, String outputImageName, @Nullable Module module) { if (objects.size() == 0) return null; // Create blank image Obj firstObject = objects.getFirst(); ImagePlus ipl = IJ.createImage(outputImageName, firstObject.getWidth(), firstObject.getHeight(), 1, 32); Image image = ImageFactory.createImage(outputImageName, ipl); // Get the image for the first object. Not adding the background to any object, // as the average background will // be added later on. Iterator iterator = objects.values().iterator(); addGaussianProfile(iterator.next(), image, true); int count = 0; int total = objects.size(); while (iterator.hasNext()) { // Adding current object's Gaussian addGaussianProfile(iterator.next(), image, true); if (module != null) module.writeProgressStatus(++count, total, "objects"); } // Determining the average background and adding this on to the image double BG = getMeanBackground(objects); ImageMath.process(image.getImagePlus(), ImageMath.CalculationModes.ADD, BG); return image; } static double getMeanBackground(Objs objects) { CumStat cs = new CumStat(); for (Obj obj : objects.values()) cs.addMeasure(obj.getMeasurement(Measurements.A_BG).getValue()); return cs.getMean(); } static void addGaussianProfile(Obj obj, Image inputImage, boolean zeroBackground) { // Create the 2D Gaussian distribution double x0 = obj.getMeasurement(Measurements.X0_PX).getValue(); double y0 = obj.getMeasurement(Measurements.Y0_PX).getValue(); double sx = obj.getMeasurement(Measurements.SIGMA_X_PX).getValue(); double sy = obj.getMeasurement(Measurements.SIGMA_Y_PX).getValue(); double A0 = obj.getMeasurement(Measurements.A_0).getValue(); double ABG = zeroBackground ? 0 : obj.getMeasurement(Measurements.A_BG).getValue(); double th = obj.getMeasurement(Measurements.THETA).getValue(); GaussianDistribution2D g2D = new GaussianDistribution2D(x0, y0, sx, sy, A0, ABG, th); // Create the blank image int minX = (int) Math.round(Math.max(0, x0 - 4 * sx)); int maxX = (int) Math.round(Math.min(x0 + 4 * sx, obj.getWidth())); int minY = (int) Math.round(Math.max(0, y0 - 4 * sy)); int maxY = (int) Math.round(Math.min(y0 + 4 * sy, obj.getHeight())); // Iterate over all pixels, evaluating the Gaussian ImageProcessor ipr = inputImage.getImagePlus().getProcessor(); for (int x = minX; x < maxX; x++) { for (int y = minY; y < maxY; y++) { float inputVal = ipr.getf(x, y); float newVal = (float) g2D.getValues(x, y)[0]; ipr.setf(x, y, inputVal + newVal); } } } @Override public Category getCategory() { return Categories.OBJECTS_PROCESS; } @Override public String getVersionNumber() { return "1.0.0"; } @Override public String getDescription() { return "Gaussian spot fitting. Can take objects as estimated locations." + "\n***Only works in 2D***" + "\n***Only works for refinement of existing spots***"; } @Override public Status process(Workspace workspace) { // Getting input image String inputImageName = parameters.getValue(INPUT_IMAGE,workspace); Image inputImage = workspace.getImage(inputImageName); ImagePlus inputImagePlus = inputImage.getImagePlus(); inputImagePlus = new Duplicator().run(inputImagePlus); // Getting input objects to refine (if selected by used) String inputObjectsName = parameters.getValue(INPUT_OBJECTS,workspace); Objs inputObjects = workspace.getObjects(inputObjectsName); // Getting parameters String sigmaMode = parameters.getValue(SIGMA_MODE,workspace); boolean limitSigma = parameters.getValue(LIMIT_SIGMA_RANGE,workspace); double minSigma = parameters.getValue(MIN_SIGMA,workspace); double maxSigma = parameters.getValue(MAX_SIGMA,workspace); boolean fixedFittingWindow = parameters.getValue(FIXED_FITTING_WINDOW,workspace); int windowWidth = parameters.getValue(WINDOW_SIZE,workspace); int maxEvaluations = parameters.getValue(MAX_EVALUATIONS,workspace); boolean removeUnfit = parameters.getValue(REMOVE_UNFIT,workspace); boolean applyVolume = parameters.getValue(APPLY_VOLUME,workspace); boolean createGaussianImage = parameters.getValue(CREATE_GAUSSIAN_IMAGE,workspace); String gaussianImageName = parameters.getValue(GAUSSIAN_IMAGE,workspace); // Running through each object, doing the fitting int count = 0; int startingNumber = inputObjects.size(); Iterator iterator = inputObjects.values().iterator(); while (iterator.hasNext()) { Obj inputObject = iterator.next(); // Getting the centroid of the current object (should be single points anyway) int x = (int) Math.round(inputObject.getXMean(true)); int y = (int) Math.round(inputObject.getYMean(true)); int z = (int) Math.round(inputObject.getZMean(true, false)); int t = inputObject.getT(); // Getting the sigma of the object int sigma; switch (sigmaMode) { case SigmaModes.FIXED_VALUE: default: sigma = (int) Math.ceil(parameters.getValue(SIGMA,workspace)); break; case SigmaModes.MEASUREMENT: double multiplier = parameters.getValue(MEASUREMENT_MULTIPLIER,workspace); sigma = (int) Math.ceil(inputObject.getMeasurement(parameters.getValue(SIGMA_MEASUREMENT,workspace)).getValue() * multiplier); break; } // Ensuring the window width is odd, then getting the half width if (windowWidth % 2 != 0) windowWidth--; int halfW = fixedFittingWindow ? windowWidth / 2 : 2*sigma; // Getting the local image region if (x - halfW < 0 || x + halfW + 1 > inputImagePlus.getWidth() || y - halfW < 0 || y + halfW + 1 > inputImagePlus.getHeight()) { assignMissingMeasurements(inputObject); continue; } // Setting limits double[][] limits = getLimits(halfW, sigma, limitSigma, minSigma, maxSigma); // Cropping image inputImagePlus.setPosition(1, z + 1, t + 1); Image preCropImage = ImageFactory.createImage("PreCrop", new ImagePlus("Slice", inputImagePlus.getProcessor())); ImageProcessor iprCrop = CropImage .cropImage(preCropImage, "Crop", x - halfW, y - halfW, halfW * 2 + 1, halfW * 2 + 1).getImagePlus() .getProcessor(); // Estimating parameters double[] pIn = estimateParameters(iprCrop, sigma); // Fitting the Gaussian and checking it reached convergence double[] pOut = GaussianFitter.fitGaussian2D(iprCrop, pIn, limits, maxEvaluations); // If the centroid has moved more than the width of the window, removing this // localisation if (pOut != null && (pOut[0] <= 1 || pOut[0] >= sigma * 2 || pOut[1] <= 1 || pOut[1] >= sigma * 2)) { pOut = null; } // If the width is outside the permitted range if ((pOut != null && limitSigma) && ((pOut[2] + pOut[3]) / 2 < sigma * minSigma || (pOut[2] + pOut[3]) / 2 > sigma * maxSigma)) { pOut = null; } // Calculating residual if (pOut == null) assignMissingMeasurements(inputObject); else assignMeasurements(inputObject, iprCrop, pOut, x, y, halfW); // If selected, any objects that weren't fit are removed if (removeUnfit & pOut == null) { inputObject.removeRelationships(); iterator.remove(); } writeProgressStatus(++count, startingNumber, "objects"); } // Adding explicit volume to spots if (applyVolume) assignVolume(inputObjects); // Creating a Gaussian image Image gaussianImage = null; if (createGaussianImage) { gaussianImage = createGaussianImage(inputObjects, gaussianImageName, this); workspace.addImage(gaussianImage); } if (showOutput) { inputObjects.showMeasurements(this, modules); if (createGaussianImage) gaussianImage.show(); // Also show output objects, if storing coordinates is somehow enabled } return Status.PASS; } @Override protected void initialiseParameters() { parameters.add(new SeparatorP(INPUT_SEPARATOR, this)); parameters.add(new InputImageP(INPUT_IMAGE, this)); parameters.add(new InputObjectsP(INPUT_OBJECTS, this)); parameters.add(new SeparatorP(FITTING_SEPARATOR, this)); parameters.add(new ChoiceP(SIGMA_MODE, this, SigmaModes.FIXED_VALUE, SigmaModes.ALL)); parameters.add(new DoubleP(SIGMA, this, 1.0)); parameters.add(new ObjectMeasurementP(SIGMA_MEASUREMENT, this)); parameters.add(new DoubleP(MEASUREMENT_MULTIPLIER, this, 1.0)); parameters.add(new BooleanP(LIMIT_SIGMA_RANGE, this, true)); parameters.add(new DoubleP(MIN_SIGMA, this, 0.25)); parameters.add(new DoubleP(MAX_SIGMA, this, 4d)); parameters.add(new BooleanP(FIXED_FITTING_WINDOW, this, false)); parameters.add(new IntegerP(WINDOW_SIZE, this, 15)); parameters.add(new IntegerP(MAX_EVALUATIONS, this, 1000)); parameters.add(new SeparatorP(OUTPUT_SEPARATOR, this)); parameters.add(new BooleanP(REMOVE_UNFIT, this, false)); parameters.add(new BooleanP(APPLY_VOLUME, this, true)); parameters.add(new BooleanP(CREATE_GAUSSIAN_IMAGE, this, false)); parameters.add(new OutputImageP(GAUSSIAN_IMAGE, this)); addParameterDescriptions(); } @Override public Parameters updateAndGetParameters() { Workspace workspace = null; Parameters returnedParameters = new Parameters(); returnedParameters.add(parameters.getParameter(INPUT_SEPARATOR)); returnedParameters.add(parameters.getParameter(INPUT_IMAGE)); returnedParameters.add(parameters.getParameter(INPUT_OBJECTS)); returnedParameters.add(parameters.getParameter(FITTING_SEPARATOR)); returnedParameters.add(parameters.getParameter(SIGMA_MODE)); if (parameters.getValue(SIGMA_MODE,workspace).equals(SigmaModes.FIXED_VALUE)) { returnedParameters.add(parameters.getParameter(SIGMA)); } else if (parameters.getValue(SIGMA_MODE,workspace).equals(SigmaModes.MEASUREMENT)) { returnedParameters.add(parameters.getParameter(SIGMA_MEASUREMENT)); String inputObjectsName = parameters.getValue(INPUT_OBJECTS,workspace); ((ObjectMeasurementP) parameters.getParameter(SIGMA_MEASUREMENT)).setObjectName(inputObjectsName); returnedParameters.add(parameters.getParameter(MEASUREMENT_MULTIPLIER)); } returnedParameters.add(parameters.getParameter(LIMIT_SIGMA_RANGE)); if ((boolean) parameters.getValue(LIMIT_SIGMA_RANGE,workspace)) { returnedParameters.add(parameters.getParameter(MIN_SIGMA)); returnedParameters.add(parameters.getParameter(MAX_SIGMA)); } returnedParameters.add(parameters.getParameter(FIXED_FITTING_WINDOW)); if ((boolean) parameters.getValue(FIXED_FITTING_WINDOW,workspace)) returnedParameters.add(parameters.getParameter(WINDOW_SIZE)); returnedParameters.add(parameters.getParameter(MAX_EVALUATIONS)); returnedParameters.add(parameters.getParameter(OUTPUT_SEPARATOR)); returnedParameters.add(parameters.getParameter(REMOVE_UNFIT)); returnedParameters.add(parameters.getParameter(APPLY_VOLUME)); returnedParameters.add(parameters.getParameter(CREATE_GAUSSIAN_IMAGE)); if ((boolean) parameters.getValue(CREATE_GAUSSIAN_IMAGE,workspace)) returnedParameters.add(parameters.getParameter(GAUSSIAN_IMAGE)); return returnedParameters; } @Override public ImageMeasurementRefs updateAndGetImageMeasurementRefs() { return null; } @Override public ObjMeasurementRefs updateAndGetObjectMeasurementRefs() { Workspace workspace = null; ObjMeasurementRefs returnedRefs = new ObjMeasurementRefs(); String inputObjectsName = parameters.getValue(INPUT_OBJECTS,workspace); ObjMeasurementRef reference = objectMeasurementRefs.getOrPut(Measurements.X0_PX); reference.setObjectsName(inputObjectsName); returnedRefs.add(reference); reference = objectMeasurementRefs.getOrPut(Measurements.Y0_PX); reference.setObjectsName(inputObjectsName); returnedRefs.add(reference); reference = objectMeasurementRefs.getOrPut(Measurements.Z0_SLICE); reference.setObjectsName(inputObjectsName); returnedRefs.add(reference); reference = objectMeasurementRefs.getOrPut(Measurements.SIGMA_X_PX); reference.setObjectsName(inputObjectsName); returnedRefs.add(reference); reference = objectMeasurementRefs.getOrPut(Measurements.SIGMA_Y_PX); reference.setObjectsName(inputObjectsName); returnedRefs.add(reference); reference = objectMeasurementRefs.getOrPut(Measurements.SIGMA_MEAN_PX); reference.setObjectsName(inputObjectsName); returnedRefs.add(reference); reference = objectMeasurementRefs.getOrPut(Measurements.X0_CAL); reference.setObjectsName(inputObjectsName); returnedRefs.add(reference); reference = objectMeasurementRefs.getOrPut(Measurements.Y0_CAL); reference.setObjectsName(inputObjectsName); returnedRefs.add(reference); reference = objectMeasurementRefs.getOrPut(Measurements.Z0_CAL); reference.setObjectsName(inputObjectsName); returnedRefs.add(reference); reference = objectMeasurementRefs.getOrPut(Measurements.SIGMA_X_CAL); reference.setObjectsName(inputObjectsName); returnedRefs.add(reference); reference = objectMeasurementRefs.getOrPut(Measurements.SIGMA_Y_CAL); reference.setObjectsName(inputObjectsName); returnedRefs.add(reference); reference = objectMeasurementRefs.getOrPut(Measurements.SIGMA_MEAN_CAL); reference.setObjectsName(inputObjectsName); returnedRefs.add(reference); reference = objectMeasurementRefs.getOrPut(Measurements.A_0); reference.setObjectsName(inputObjectsName); returnedRefs.add(reference); reference = objectMeasurementRefs.getOrPut(Measurements.A_BG); reference.setObjectsName(inputObjectsName); returnedRefs.add(reference); reference = objectMeasurementRefs.getOrPut(Measurements.THETA); reference.setObjectsName(inputObjectsName); returnedRefs.add(reference); reference = objectMeasurementRefs.getOrPut(Measurements.ELLIPTICITY); reference.setObjectsName(inputObjectsName); returnedRefs.add(reference); reference = objectMeasurementRefs.getOrPut(Measurements.RESIDUAL); reference.setObjectsName(inputObjectsName); returnedRefs.add(reference); return returnedRefs; } @Override public ObjMetadataRefs updateAndGetObjectMetadataRefs() { return null; } @Override public MetadataRefs updateAndGetMetadataReferences() { return null; } @Override public ParentChildRefs updateAndGetParentChildRefs() { return null; } @Override public PartnerRefs updateAndGetPartnerRefs() { return null; } @Override public boolean verify() { return true; } void addParameterDescriptions() { parameters.get(INPUT_IMAGE) .setDescription("Image from the workspace to which the 2D Gaussian profiles will be fit."); parameters.get(INPUT_OBJECTS).setDescription( "Objects for which 2D Gaussians will be fit. A single Gaussian will be calculated for each object, with the centroid of the input object used as the starting estimate for the Gaussian centroid. Fit parameters will be stored as measurements associated with the relevant input objects."); parameters.get(SIGMA_MODE).setDescription( "Controls how the initial estimate of Gaussian sigma will be calculated. Note: The initial radius value can also influence the range of accepted sigma values (see \"" + LIMIT_SIGMA_RANGE + "\" parameter).
    " + "
  • \"" + SigmaModes.FIXED_VALUE + "\" The same initial sigma value is applied to the fitting of all objects. This value is provided by the \"" + SIGMA + "\" parameter.
  • " + "
  • \"" + SigmaModes.MEASUREMENT + "\" A different sigma value is applied to the fitting of each object. This value is provided by the measurement (specified by \"" + SIGMA_MEASUREMENT + "\") associated with the object being fit.
"); parameters.get(SIGMA).setDescription("If \"" + SIGMA_MODE + "\" is set to \"" + SigmaModes.FIXED_VALUE + "\", this is the initial sigma value that will be used in the fitting of all objects."); parameters.get(SIGMA_MEASUREMENT).setDescription("If \"" + SIGMA_MODE + "\" is set to \"" + SigmaModes.MEASUREMENT + "\", this is the measurement that will provide the intial sigma value used in the fitting of each object."); parameters.get(MEASUREMENT_MULTIPLIER).setDescription("If using object-associated measurements as the intial estimate of sigma, these measurements can be systematically increased or decreased with this multiplier. Values less than 1 will reduce the sigma estimate, while values greater than 1 will increase it. To use the default sigma value, set this to 1."); parameters.get(LIMIT_SIGMA_RANGE).setDescription("When selected, the final sigma value must lie within a specific range. This range is controlled by the \""+MIN_SIGMA+"\" and \""+MAX_SIGMA+"\" parameters."); parameters.get(MIN_SIGMA).setDescription("If limiting the final sigma range (\""+LIMIT_SIGMA_RANGE+"\" parameter selected), this value controls the minimum allowed sigma value. The minimum range is specified as a multiplier of the input sigma estimate. For example, an estimate sigma value of 4 with a \""+MIN_SIGMA+"\" multiplier of 0.5 forces the predicted sigma to have a value of at least 2. Variable sigma ranges allow measurement-based sigma estimates to have appropriate limits."); parameters.get(MAX_SIGMA).setDescription("If limiting the final sigma range (\""+LIMIT_SIGMA_RANGE+"\" parameter selected), this value controls the maximum allowed sigma value. The maximum range is specified as a multiplier of the input sigma estimate. For example, an estimate sigma value of 4 with a \""+MAX_SIGMA+"\" multiplier of 3 forces the predicted sigma to have a value of at most 12. Variable sigma ranges allow measurement-based sigma estimates to have appropriate limits."); parameters.get(FIXED_FITTING_WINDOW).setDescription("When selected, the size of the cropped image region, to which the 2D Gaussian is fit, is controlled by the \""+WINDOW_SIZE+"\" parameter. Otherwise, the window size is automatically selected as 4 times the initial estimate of sigma, plus 1."); parameters.get(WINDOW_SIZE).setDescription("If using a fixed fitting window (\""+FIXED_FITTING_WINDOW+"\" parameter selected), this is the size of the image crop (centered on the object being fit) that will be used."); parameters.get(MAX_EVALUATIONS).setDescription("The maximum number of iterations the fitting algorithm can use."); parameters.get(REMOVE_UNFIT).setDescription("If selected, any objects for which fits can't be determined (fitter runs out of evaluations before completing) will be removed from the input object set."); parameters.get(APPLY_VOLUME).setDescription("If selected, the coordinates of the input objects will be updated to be circles. The output circles will be centred on the best-fit location and have radii equal to the x-axis sigma value."); parameters.get(CREATE_GAUSSIAN_IMAGE).setDescription("If selected, an image containing all calculated Gaussian fits will be added to the workspace."); parameters.get(GAUSSIAN_IMAGE).setDescription("If \""+CREATE_GAUSSIAN_IMAGE+"\" is selected, this is the name of the output Gaussian profile image that will be added to the workspace."); } } // when signax_0:sigmay_o is >1.5, delete spot... i - 1 ;




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