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Colocalization analysis in fluorescence micrographs: verification of a more accurate calculation of Pearson's correlation coefficient

(2010) MICROSCOPY AND MICROANALYSIS. 16(6). p.710-724
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Abstract
One of the most routine uses of fluorescence microscopy is colocalization, i.e., the demonstration of a relationship between pairs of biological molecules. Frequently this is presented simplistically by the use of overlays of red and green images, with areas of yellow indicating colocalization of the molecules. Colocalization data are rarely quantified and can be misleading. Our results from both synthetic and biological datasets demonstrate that the generation of Pearson's correlation coefficient between pairs of images can overestimate positive correlation and fail to demonstrate negative correlation. We have demonstrated that the calculation of a thresholded Pearson's correlation coefficient using only intensity values over a determined threshold in both channels produces numerical values that more accurately describe both synthetic datasets and biological examples. Its use will bring clarity and accuracy to colocalization studies using fluorescent microscopy.
Keywords
Pearson's correlation, algorithm, fluorescence microscopy, colocalization, quantitation, IMAGE ACQUISITION, MICROSCOPY, CELLS, RECEPTORS

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Citation

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Chicago
Barlow, Andrew L, Alasdair MacLeod, Samuel Noppen, Jeremy Sanderson, and Chris Guerin. 2010. “Colocalization Analysis in Fluorescence Micrographs: Verification of a More Accurate Calculation of Pearson’s Correlation Coefficient.” Microscopy and Microanalysis 16 (6): 710–724.
APA
Barlow, A. L., MacLeod, A., Noppen, S., Sanderson, J., & Guerin, C. (2010). Colocalization analysis in fluorescence micrographs: verification of a more accurate calculation of Pearson’s correlation coefficient. MICROSCOPY AND MICROANALYSIS, 16(6), 710–724.
Vancouver
1.
Barlow AL, MacLeod A, Noppen S, Sanderson J, Guerin C. Colocalization analysis in fluorescence micrographs: verification of a more accurate calculation of Pearson’s correlation coefficient. MICROSCOPY AND MICROANALYSIS. 2010;16(6):710–24.
MLA
Barlow, Andrew L, Alasdair MacLeod, Samuel Noppen, et al. “Colocalization Analysis in Fluorescence Micrographs: Verification of a More Accurate Calculation of Pearson’s Correlation Coefficient.” MICROSCOPY AND MICROANALYSIS 16.6 (2010): 710–724. Print.
@article{1089037,
  abstract     = {One of the most routine uses of fluorescence microscopy is colocalization, i.e., the demonstration of a relationship between pairs of biological molecules. Frequently this is presented simplistically by the use of overlays of red and green images, with areas of yellow indicating colocalization of the molecules. Colocalization data are rarely quantified and can be misleading. Our results from both synthetic and biological datasets demonstrate that the generation of Pearson's correlation coefficient between pairs of images can overestimate positive correlation and fail to demonstrate negative correlation. We have demonstrated that the calculation of a thresholded Pearson's correlation coefficient using only intensity values over a determined threshold in both channels produces numerical values that more accurately describe both synthetic datasets and biological examples. Its use will bring clarity and accuracy to colocalization studies using fluorescent microscopy.},
  author       = {Barlow, Andrew L and MacLeod, Alasdair and Noppen, Samuel and Sanderson, Jeremy and Guerin, Chris},
  issn         = {1431-9276},
  journal      = {MICROSCOPY AND MICROANALYSIS},
  keyword      = {Pearson's correlation,algorithm,fluorescence microscopy,colocalization,quantitation,IMAGE ACQUISITION,MICROSCOPY,CELLS,RECEPTORS},
  language     = {eng},
  number       = {6},
  pages        = {710--724},
  title        = {Colocalization analysis in fluorescence micrographs: verification of a more accurate calculation of Pearson's correlation coefficient},
  url          = {http://dx.doi.org/10.1017/S143192761009389X},
  volume       = {16},
  year         = {2010},
}

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