Fourier ring correlation was recently proposed in fluorescence microscopy as an objective measure of image resolution. In this study we show that FRC has serious limitations. For several common image processing techniques, we show that FRC can be made to be systematically biased to classify objectively lower resolution images as better resolved than objectively higher resolution images. Thus, rather than providing an objective, realistic measure of spatial resolution, care must be taken in interpreting FRC-based results. In revealing some of FRC’s limitations, this study also highlights the need for a more objective, principled approach to estimating resolution.
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