# Jeff Hardin, Dept. of Integrative B )logy # Univ. of Wisconsin-Madison # jdhardin@wisc.edu # July 30, 2025 #Additional colors could be added via org.jfree.chart.ChartColor #Green = #32a852 #Purple = #7132a8 #Dark red = #9C2005 #Dark brown = #754F04 #Light brown = #9C6905 #This script imports data from one or more CSV files in the form x[i],y[i] as paired columns #using ImageJ's built-in support for importing CSV files in a ResultsTable #and ImageJ's built-in Plot functions. #Currently uses the hard-wired colors built into ImageJ's Plot class and a few hexadecimal colors. #Use the PlotWindow "More >> " button to change range, line thickness, colors, etc. #Using JFreeChart would allow export as an editable SVG file, but has less flexibiity #in changing appearance after generating the graph from org.jfree.data.statistics import Statistics from javax.swing import JFrame import java.awt.Color as Color from java.awt import Dimension, BasicStroke import java.awt.Frame as Frame import java.awt.Window as Window from java.io import File as File from java.lang import System as System from ij import WindowManager as WindowManager from ij.plugin.frame import RoiManager as RoiManager from ij.process import ImageStatistics as ImageStatistics from ij.measure import Measurements as Measurements from ij import IJ as IJ from ij.measure import CurveFitter as CurveFitter from ij.gui import Plot as Plot from ij.gui import PlotWindow as PlotWindow from ij.gui import ImageWindow as ImageWindow from ij.text import TextWindow from ij.gui import GenericDialog from ij import ImagePlus as ImagePlus from ij.io import FileInfo as FileInfo from ij.measure import ResultsTable as ResultsTable from ij import WindowManager as WindowManager import math import os from os import path, mkdir import csv #below from #https://stackoverflow.com/questions/736043/checking-if-a-string-can-be-converted-to-float-in-python def isfloat(value): try: float(value) return True except ValueError: return False def doPlot(): k = len(listOfPaths) if (k < 2): IJ.showMessage("Incorrect data structure","This script required more than one dataset.") return None #Ask for options gd = GenericDialog("Options") gd.addCheckbox("Combine plots", True) gd.addCheckbox("Plot Y mean values", True) gd.addCheckbox("Show error bars", True) items = ["Std err", "Std dev"]; gd.addRadioButtonGroup("Error Bars", items, 1, 2, "Std err"); gd.addCheckbox("Curve fit means", True) gd.addChoice("Curve fitting method:", ["Single", "Double"], "Single") gd.addCheckbox("Force curve fit through origin for single exponential", True) gd.showDialog() if gd.wasOKed(): combinePlots = gd.getNextBoolean() plotMeans = gd.getNextBoolean() plotStdErrs = gd.getNextBoolean() errorBarType = gd.getNextRadioButton() fitMeans = gd.getNextBoolean() fitMethod = gd.getNextChoice() throughOrigin = gd.getNextBoolean() else: return None lineseparator = "\n" cellseparator = "," legendString = "" myTable = ResultsTable() #set the next to True so that we can throw NaN cells later myTable.setNaNEmptyCells(True) if (combinePlots): myPlot=Plot("Combined", "X", "Y") myPlot.setLineWidth(2) xMin = float() xMax = float() xMax = 0 xMin = 0 plotColor=[] plotColor = ("black", "red", "blue", "#32a852", "#7132a8", "#9C2005", "#754F04", "magenta", "darkGray", "gray", "lightGray", "green", "#9C6905", "cyan", "orange", "pink", "yellow"); #Call below is a feature of Fiji/IJ2 #See https://imagej.net/scripting/parameters #for i in range(0, k): #print(listOfPaths[i]) for i in range(0, k): #read in FRAP curve data from CSV file #copies the whole file to an array of lines #have to type cast pathnames to Python str text_file = open(str(listOfPaths[i]), "r") #read whole file to a string data = text_file.read() #close file text_file.close() #separate into lines of text lines=data.split(lineseparator) numRows = len(lines) # recreates the columns headers labels=lines[0].split(cellseparator) #get total columns numCol = len(labels) #declare arrays to hold data points x1=[] y1=[] #IJ.open(str(listOfPaths[i])) with open(str(listOfPaths[i]), 'r') as read_obj: # pass the file object to reader() to get the reader object csv_reader = csv.reader(read_obj) # Iterate over each row in the csv using reader object header = next(csv_reader) for row in csv_reader: #need to add code to check for empty cells, since ImageJ #produces CSV files in which there can be unequal numbers of rows with empty cells #Jython interpreter crashes when trying to convert to float if cell is blank #ImageJ plot functions are savvy about blank cells, but this code isn't! if (isfloat(row[0])): x1.append(float(row[0])) if (isfloat(row[1])): y1.append(float(row[1])) #get min and mx for X axis if (min(x1) < xMin): xMin = min(x1) if (max(x1) > xMax): xMax = max(x1) #IJ.log("x1 length:") #IJ.log(str(len(x1))) #IJ.log("y1 length:") #IJ.log(str(len(y1))) #Use a Java method instead (need JVM 7+) file = File(str(listOfPaths[i])) #get file name using getName() filename = file.getName() #Now let's plot the XY data using ImageJ's Plot function if (combinePlots): if (i < len(plotColor)): colorString = plotColor[i] else: colorString = "black" myPlot.setColor(colorString) myPlot.setLineWidth(2.0) #Removes gridlines; delete if you like these... myPlot.setFormatFlags(0x330f) myPlot.add("Line",x1,y1) if (i == 0): legendString = legendString + File.getName(listOfPaths[i]) else: legendString = legendString + "\t" + File.getName(listOfPaths[i]) #create ResultTable for data for m in range(len(x1)): #IJ.log("m:") #IJ.log(str(m)) #if first dataset, create new rows if (i == 0 ): myTable.incrementCounter() myTable.addValue(filename + "-x",x1[m]) myTable.addValue(filename + "-y",y1[m]) else: #myTable.setValue(filename + "-x",m,x1[m]) myTable.setValue(filename + "-y",m,y1[m]) #Calculate stats and graph mean + std err if (len(listOfPaths) > 1): yValues = [] yMeans = [] yStdDevs = [] yMean = 0 for j in range(len(x1)): for i in range(0, len(listOfPaths)): if (j == 0): yValues.append(myTable.getValueAsDouble(i+1,j)) else: yValues[i] = myTable.getValueAsDouble(i+1,j) #Add columns for stats #print(yValues) myTable.setValue("Mean",j,Statistics.calculateMean(yValues)) #Still need to add code to check for NaN if n = 2 myTable.setValue("Std Dev",j,Statistics.getStdDev(yValues)) myTable.setValue("Std Err",j,Statistics.getStdDev(yValues)/math.sqrt(len(listOfPaths))) myTable.show("Data") if (combinePlots): myPlot.show() myPlot.setLegend(legendString,Plot.AUTO_POSITION) myPlot.setLimitsToFit(True) #now graph the mean and std err if (plotMeans): #IJ.log("length of x1:") #IJ.log(str(len(x1))) myMeanPlot = Plot("Mean value", "X", "Y mean") myMeanPlot.setLineWidth(2) myMeanPlot.setColor("red") #Removes gridlines; delete if you like these... myMeanPlot.setFormatFlags(0x330f) #couldn't get myPlot.getColumn("Mean") to work, so do it manually ymean = [] for i in range(len(x1)): ymean.append(myTable.getValue("Mean",i)) myMeanPlot.add("Line",x1,ymean) legendString = "X vs. mean" if (plotStdErrs): if (errorBarType == "Std err"): legendString = "X vs. mean +/- SEM" yStdErr = [] for i in range(len(x1)): yStdErr.append(myTable.getValue("Std Err",i)) myMeanPlot.add("error bars", yStdErr) else: legendString = "X vs. mean +/- SD" yStdDev = [] for i in range(len(x1)): yStdDev.append(myTable.getValue("Std Dev",i)) myMeanPlot.add("error bars", yStdDev) myMeanPlot.setLimitsToFit(True) myMeanPlot.setColor("black") myMeanPlot.setLegend(legendString,Plot.AUTO_POSITION) myMeanPlot.show() if (fitMeans): # Fitter yFit = [] fitter = CurveFitter(x1, ymean) if fitMethod == "Single": if throughOrigin: fitter.doFit(CurveFitter.EXP_RECOVERY_NOOFFSET) else: fitter.doFit(CurveFitter.EXP_RECOVERY) #myMeanPlot.add("fit", yStdErr) #fitter.plot param_values = fitter.getParams() for i in range(len(x1)): yFit.append( fitter.f( fitter.getParams(), x1[i]) ) myTable.setValue("Fit",i,yFit[i]) myMeanPlot.setColor("blue") myMeanPlot.add("Line",x1,yFit) myMeanPlot.setColor("black") myMeanPlot.setLegend(legendString + "\nFit",Plot.AUTO_POSITION) myMeanPlot.show() myTable.show("Data") else: eqn = "y = a*(1-exp(-b*x)) +c*(1-exp(-d*x)) + e" params = fitter.doCustomFit(eqn, None, False) for i in range(len(x1)): yFit.append( fitter.f( fitter.getParams(), x1[i]) ) myTable.setValue("Fit",i,yFit[i]) myMeanPlot.setColor("blue") myMeanPlot.add("Line",x1,yFit) myMeanPlot.setColor("black") myMeanPlot.setLegend(legendString + "\nFit",Plot.AUTO_POSITION) myMeanPlot.show() myTable.show("Data") #Output FRAP paramaters #need to fix this! time_units = "sec" myTextWindow = TextWindow("FRAP Results","",600,300) myTextWindow.append("Fit FRAP curve by " + fitter.getFormula() ) param_values = fitter.getParams() if fitMethod == "Single": if throughOrigin: myTextWindow.append("Fit constrained through origin: TRUE") else: myTextWindow.append("Fit constrained through origin: FALSE") thalf = math.log(2) / param_values[1] mobile_fraction = param_values[0] str1 = ('Half-life = %.2f ' + time_units) % thalf myTextWindow.append( str1 ) str2 = "Mobile fraction = %.1f %%" % (100 * mobile_fraction) myTextWindow.append( str2 ) myTextWindow.append( "" ) myTextWindow.append("*******Details********" + fitter.getResultString() ) else: thalf1 = math.log(2) / param_values[1] thalf2 = math.log(2) / param_values[3] mobile_fraction = param_values[0] + param_values[2] str1 = ('Half-life #1= %.2f ' + time_units) % thalf1 myTextWindow.append( str1 ) str2 = ('Half-life #2= %.2f ' + time_units) % thalf2 myTextWindow.append( str2 ) str3 = "Mobile fraction = %.1f %%" % (100 * mobile_fraction) myTextWindow.append( str3 ) myTextWindow.append( "" ) myTextWindow.append("*******Details********" + fitter.getResultString() ) #main #@ File[] listOfPaths (label="select files", style="files") doPlot()