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A simple image correction method for high-throughput microscopy

Nature Methods volume 11, page 602 (2014)Cite this article

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Advances in high-throughput fluorescence microscopy (HTFM) and image informatics have allowed single-cell phenotypic variability to be characterized and related to putative biological functions. However, estimates of single-cell variability may be skewed by inaccurate image correction. Furthermore, methods used in the HTFM literature do not generally take advantage of shared image properties, instead correcting each image independently. Here, we revisit the basic mathematical framework underlying image correction to show how the true distributions of commonly measured single-cell features are altered in uncorrected images. We also report an observation that allows the use of a simple method for accurate image correction in HTFM.

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Figure 1: A simple method for image correction in HTFM to improve estimates of single-cell phenotypes.

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Acknowledgements

We thank J. Shay (University of Texas Southwestern Medical Center) for kindly providing human colonic epithelial cells. This work was supported by US National Cancer Institute R01CA133253 (S.J.A.); US National Institutes of Health R01 GM071794 (S.J.A.), R01 GM081549 (L.F.W.) and T32 GM008203 (A.D.C.); the Welch Foundation I-1619 (S.J.A.) and I-1644 (L.F.W.) and the Howard Hughes Medical Institute Med into Grad Initiative (A.D.C.).

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Authors and Affiliations

  1. Green Center for Systems Biology, University of Texas Southwestern Medical Center, Dallas, Texas, USA

    Adam D Coster, Chonlarat Wichaidit, Satwik Rajaram, Steven J Altschuler & Lani F Wu

Authors
  1. Adam D Coster
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  2. Chonlarat Wichaidit
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  3. Satwik Rajaram
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  4. Steven J Altschuler
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  5. Lani F Wu
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Corresponding authors

Correspondence to Steven J Altschuler or Lani F Wu.

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The authors declare no competing financial interests.

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Coster, A., Wichaidit, C., Rajaram, S. et al. A simple image correction method for high-throughput microscopy. Nat Methods 11, 602 (2014). https://doi.org/10.1038/nmeth.2971

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