Analysis of Image Similarity and Relationship



The ability of fluorescence microscopy to simultaneously image multiple specific molecules of interest has allowed biologists to infer macromolecular organization and colocalization in fixed and live samples. However, a number of factors could affect these analyses, and colocalization is a misnomer. We propose that image similarity coefficient as a better and more descriptive term. In this chapter we will discuss many of the factors involved with determining image similarity including our perception of color in images. In addition, the correct use of several commonly accepted methods such as Pearson’s correlation coefficient, Manders’ overlap coefficient, and Spearman’s ranked correlation coefficient is discussed.


Colocalization Manders’ coefficient Pearson’s coefficient Spearman’s ranked correlation coefficient Dynamic range 


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© Springer Nature Switzerland AG 2018

Authors and Affiliations

  1. 1.Advanced Imaging CenterHoward Hughes Medical Institute Janelia Research CampusAshburnUSA

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