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Histochem Cell Biol. 2014 Jun;141(6):605-12. doi: 10.1007/s00418-014-1208-z. Epub 2014 Mar 19.

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

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Centre for Vascular Research and Australian Centre for NanoMedicine, University of New South Wales, Sydney, Australia.


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.

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