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. Author manuscript; available in PMC: 2018 Feb 21.
Published in final edited form as: Methods Mol Biol. 2016;1389:111–126. doi: 10.1007/978-1-4939-3302-0_7

Figure 4. Setting the Spot Mask.

Figure 4

To accurately design a spot mask first the ‘bright’ option must be checked. This ensures the mask obtains the bright regions from an image regardless of any intensity differences from one spot to another. Next, alter the spot-to background ratio and the radius of the spot mask to ensure the mask covers the spots but does not include any background.