Techniques

A universal analysis tool for the detection of asymmetric signal distribution in microscopic images†

  1. Maja Matis1,*,
  2. Jeffrey D. Axelrod1,
  3. Milos Galic2,*

Article first published online: 23 JUN 2012

DOI: 10.1002/dvdy.23818

Issue

Developmental Dynamics

Developmental Dynamics

Volume 241, Issue 8, pages 1301–1309, August 2012

Additional Information

How to Cite

Matis, M., Axelrod, J. D. and Galic, M. (2012), A universal analysis tool for the detection of asymmetric signal distribution in microscopic images. Dev. Dyn., 241: 1301–1309. doi: 10.1002/dvdy.23818

Author Information

  1. 1

    Department of Pathology, Stanford University School of Medicine, Stanford, California

  2. 2

    Chemical and Systems Biology, Stanford University School of Medicine, Stanford, California

*Department of Pathology, Stanford University School of Medicine, 300 Pasteur Drive, L235, Stanford, CA 94305

  1. †

    M.M. performed the experiments and M.G. made the analysis. M.M, J.D.A., and M.G. discussed all results and wrote the report.

Publication History

  1. Issue published online: 17 JUL 2012
  2. Article first published online: 23 JUN 2012
  3. Accepted manuscript online: 11 JUN 2012 10:51AM EST
  4. Manuscript Accepted: 31 MAY 2012

Funded by

  • AXA
  • Swiss National Science Foundation. Grant Number: PBBSP3-123159
  • Novartis Jubilaeumsstiftung
  • NIH. Grant Numbers: R01 GM059823, R01 GM098582

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Keywords:

  • fluorescent microscopy;
  • asymmetric protein and organelle localization;
  • image analysis;
  • cross-correlation;
  • PCP;
  • planar cell polarity

Abstract

Background: Polarization of tissue is achieved by asymmetric distribution of proteins and organelles within individual cells. However, existing quantitative assays to measure this asymmetry in an automated and unbiased manner suffer from significant limitations. Results: Here, we report a new way to assess protein and organelle localization in tissue based on correlative fluorescence analysis. As a proof of principle, we successfully characterized planar cell polarity dependent asymmetry in developing Drosophila melanogaster tissues on the single cell level using fluorescence cross-correlation. Conclusions: Systematic modulation of signal strength and distribution show that fluorescence cross-correlation reliably detects asymmetry over a broad parameter space. The novel method described here produces robust, rapid, and unbiased measurement of biometrical properties of cell components in live tissue that is readily applicable in other model systems. Developmental Dynamics 241:1301–1309, 2012. © 2012 Wiley Periodicals, Inc.

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