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foreign we will use image J software to remove these Speckles from the image this is to achieve an accurate quantification such as counting the number of dappy stain cells or performing a fluorescence intensity measurement the first step is to convert the image into an 8-bit image adjust the threshold in the image such that the foreground is completely red in color with a dark background at times the Speckles may be undistinguished from the cells and may need a manual separation select the Color Picker from above and click on the black color now use the brush tool to draw a line separating the Speckles from the dappy staining once the parameters are set click on apply to exclude the Speckles click on analyze and analyze particles in the analyze particles window select either ins or pixels as the unit of measurement set the size such that the speckles are excluded during the analysis in the show option select mask these options can be left unchecked once completed click ok now the speck
