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

Andrew L Barlow, Alasdair MacLeod, Samuel Noppen UGent, Jeremy Sanderson UGent and Chris Guerin UGent (2010) MICROSCOPY AND MICROANALYSIS. 16(6). p.710-724
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.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
Pearson's correlation, algorithm, fluorescence microscopy, colocalization, quantitation, IMAGE ACQUISITION, MICROSCOPY, CELLS, RECEPTORS
journal title
MICROSCOPY AND MICROANALYSIS
Microsc. microanal.
volume
16
issue
6
pages
710 - 724
Web of Science type
Article
Web of Science id
000284715300008
JCR category
MATERIALS SCIENCE, MULTIDISCIPLINARY
JCR impact factor
2.179 (2010)
JCR rank
47/219 (2010)
JCR quartile
1 (2010)
ISSN
1431-9276
DOI
10.1017/S143192761009389X
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
1089037
handle
http://hdl.handle.net/1854/LU-1089037
date created
2010-12-17 13:06:28
date last changed
2016-12-19 15:46:43
@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},
}

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.