![]() ![]() Get the filename from the title of the image that's open for adding to the results table // We do this instead of using the imageFile parameter so that the // directory path is not included on the table filename = getTitle() Store the number of results before executing the commands, // so we can add the filename just to the new results prevNumResults = nResults Save the results data saveAs( "results", outputFile) SetBatchMode( false) // Now disable BatchMode since we are finished updateResults() // Update the results table so it shows the filenames // Show a dialog to allow user to save the results file outputFile = File. Get the list of files from that directory // NOTE: if there are non-image files in this directory, it may cause the macro to crash fileList = getFileList( inputDirectory) Show the user a dialog to select a directory of images inputDirectory = getDirectory( "Choose a Directory of Images") Macro to measure Area, Intensity, Perimeter, and Shape of directory of images run( "Clear Results") // clear the results table of any previous measurements // The next line prevents ImageJ from showing the processing steps during // processing of a large number of images, speeding up the macro setBatchMode( true) We will call the function “processImage” and it will contain the lines that were recorded using the macro recorder. The Macro code box will pop up.įirst, we will create a function to execute the steps we just recorded. To create a new macro, on the ImageJ menu, click Plugins > New > Macro. The second line actually calls the Analyze Particles command and gives it the parameters needed to execute the function, this is the line we need. The first line where this appears is not necessary, since that line will show the dialog box for the Analyze Particles window, so you can delete that line, since for automating the analysis of a whole plate, we don’t want to show the Analyze Particles dialog for every image. Also notice that there are 2 lines which show the run(“Analyze Particles…”) command. The first line is commented out (everything on a line “//” in ImageJ code is ignored, known as a comment), since that line would show the Threshold dialog box, and we want to set the threshold automatically, not show the dialog box for user input. The results table will pop up, showing you 4 rows, one row for each cell detected. Select “Display results” since we want to save the results to the results table after analysis. We enter 1 um² as the lower limit, that way we will eliminate any specks of dust that show up in the image. For the size parameter, enter in 1.00-Infinity. The Analyze Particles dialog will pop up. Click the Analyze menu, and then select Analyze Particles. The next step is to use the Analyze Particles function in ImageJ to detect the cells and measure their characteristics. Also select Limit to threshold, otherwise ImageJ will calculate this measurements on the whole image instead of the thresholded areas, which is not what we want to do. Select Area, Mean gray value, Perimeter, and Shape descriptors (for circularity measurement). On the ImageJ menu, select Analyze > Set Measurements. ![]() Next we need to select the measurements we want to obtain from the particles. ![]() Comparison to Traditional Plant Tissue Clearing.Immune Cell Population Analysis for Immuno-oncology.Biomarker Quantification and Colocalization.Highly-Multiplexed Immunohistochemistry Services.Cleared Tissue 3D Imaging and Analysis Services.Antibody Penetration and Pharmacokinetics.In Vitro and 3D Cell Culture Assay Services. ![]() Does anyone know of a solution within ImageJ, or alternatively another method such as through MATLAB?Įxample of two neurons, green circles indicate ROI position from day one scan, while current neuron position is offset to the right. This is necessary as I am generally dealing with hundreds of ROIs at once, making manually adjusting each ROI prohibitive. While Multi Measure does have a tool to update the position of individual ROIs, I cannot find (in ImageJ or on the internet) a way to shift all of my ROIs at once in the same direction. This means my ROI map from the first day does not line up with where the neurons are in the second day scan. Between days, my scans tend to be off from one another slightly in the XY coordinates. The plugin has been working wonderfully for this, however in more recent experiments I am repeatedly imaging from the same area over multiple days. I’ve been using the Multi Measure plugin to select ROIs and get the mean intensity of each ROI at each time point in the stack. My experiments involve two photon calcium imaging of neurons in vivo, which outputs data as a large time series of individual images (stack). ![]()
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