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This tutorial demonstrates two methods for dot blood quantification: the percent area method and the integrated density method. To use the percent area method, open the dot plot image in ImageJ and invert it to display white dots on a dark background. Use the selection tools to mark background noise, then click "Analyze" and "Measure" to find the main intensity (e.g., 54.75). Deselect the background and go to "Process," then "Math," and "Subtract" to remove background noise using the mean intensity value. Ensure the image has a dark foreground and white background for quantification by inverting the image again and using the rectangular selection tool to enclose all dots. Finally, click on "Analyze," select "Gels," and then "First Lane."
