Histochemistry and Cell Biology

, Volume 141, Issue 6, pp 605–612 | Cite as

Method for co-cluster analysis in multichannel single-molecule localisation data

Original Paper

Abstract

We demonstrate a combined univariate and bivariate Getis and Franklin’s local point pattern analysis method to investigate the co-clustering of membrane proteins in two-dimensional single-molecule localisation data. This method assesses the degree of clustering of each molecule relative to its own species and relative to a second species. Using simulated data, we show that this approach can quantify the degree of cluster overlap in multichannel point patterns. The method is validated using photo-activated localisation microscopy and direct stochastic optical reconstruction microscopy data of the proteins Lck and CD45 at the T cell immunological synapse. Analysing co-clustering in this manner is generalizable to higher numbers of fluorescent species and to three-dimensional or live cell data sets.

Keywords

Cluster analysis Super-resolution Co-localisation PALM STORM 

Notes

Acknowledgments

D.M.O. is supported by a Marie Curie Career Integration Grant (CIG) Ref 334303. KG is supported by the National Health and Medical Research Council of Australia and the Australian Research Council.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  1. 1.Centre for Vascular Research and Australian Centre for NanoMedicineUniversity of New South WalesSydneyAustralia
  2. 2.Department of MathematicsImperial College LondonLondonUK
  3. 3.Department of Physics and Randall Division of Cell and Molecular BiophysicsKing’s College LondonLondonUK

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