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We develop image analysis software to get quantitative and objective answers to our questions. We have developed and patented the method RBNCC (replicate based noise corrected correlation) where image noise, which is unavoidable and leads to the underestimation of the underlying correlation, can be eliminated from correlation measurements. RBNCC forms the basis for our company No More Noise.
We have performed detailed studies on coefficients developed for use in colocalisation analyses revealing that several are not fit for their purpose. We advocate that coloocalisation analysis should be divided into the two subgroups co-occurrence and correlation and that only pixels where both fluorophores are present should be included in correlation analyses. We now investigate how deconvolution affects correlation analysis and how different intracellular distributions of molecules are represented by our division of colocalisation analysis into co-occurrence and correlation.
Adler, J, Parmryd, I. (2013) Colocalization analysis in fluorescence microscopy. Methods Mol. Biol. 931, 97-109
Adler J, Parmryd I. (2014) Quantifying Colocalisation: empty pixels, thresholding and the Hcoef. PLoS One 9:111983