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Statistical tests for measures of colocalization in biological microscopy
Abstract
Summary: Colocalization analysis is the most common technique used for quantitative analysis of fluorescence microscopy images. Several metrics have been developed for measuring the colocalization of two probes, including Pearson's correlation coefficient (PCC) and Manders’ correlation coefficient (MCC). However, once measured, the meaning of these measurements can be unclear; interpreting PCC or MCC values requires the ability to evaluate the significance of a particular measurement, or the significance of the difference between two sets of measurements. In previous work, we showed how spatial autocorrelation confounds randomization techniques commonly used for statistical analysis of colocalization data. Here we use computer simulations of biological images to show that the Student's one‐sample t‐test can be used to test the significance of PCC or MCC measurements of colocalization, and the Student's two‐sample t‐test can be used to test the significance of the difference between measurements obtained under different experimental conditions.
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Initial attempts to quantify colocalization by imaging in two or three dimensions followed by application of Pearson's or Manders' coefficients did not accurately reflect the degree of overlap observed, likely because of the significant degree of cytoplasmic EHD1 haze that interferes with the analysis.
“did not accurately reflect the degree of overlap observed, likely because of the significant degree of cytoplasmic EHD1 haze that interferes with the analysis, especially in cells imaged without LRP1 internalization.”
From Sorting nexin 17 (SNX17) links endosomal sorting to Eps15 homology domain protein 1 (EHD1)–mediated fission machinery (2020) · cites this paper as [49]
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Metadata
- Authors
- MCDONALD, J. H., & DUNN, K. W.
- Affiliations
- Department of Biological Sciences University of Delaware Newark Delaware U.S.A.
- Department of Medicine Division of Nephrology Indiana University Medical Center Indianapolis Indiana U.S.A.
- Venue
- Journal of Microscopy
- Publisher
- Wiley
- Published
- 2013-10-10
- Volume
- 252
- Issue
- 3
- Pages
- 295-302
- ISSN
- 0022-2720
- 1365-2818
- PMC
- PMC4428547
- Cited by
- 140
- References
- 0
- Language
- en
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