def main(): # Open a .ome.tif image from the Flexoscope. impath = IJ.getFilePath("Choose .ome.tiff file") channels = Opener.openUsingBioFormats(impath) # Show image # imp.show() # straight to channels object sames memory. # Split channels. channels = ChannelSplitter().split(channels) # Process channel 1. # subtractzproject(imp, projectionMethod="Median") channels[1] = ImagePlus() channels.append(ImagePlus()) channels[1] = subtractzproject(channels[0]) IJ.run(channels[0], "Enhance Contrast...", "saturated=0.3 normalize process_all use") IJ.run(channels[0], "8-bit", "") IJ.run(channels[1], "Square", "stack") IJ.run(channels[1], "Enhance Contrast...", "saturated=0.3 normalize process_all use") IJ.run(channels[1], "8-bit", "") # Merge channels. merge = RGBStackMerge().mergeChannels(channels, True) # boolean keep merge.show()