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Reply To: Molecular Brightness analysis of GPCR oligomerization in the presence of spatial heterogeneity

Biener, G.; Stoneman, M.; Raicu, V.

2019-11-11 biophysics
10.1101/822296 bioRxiv
Show abstract

Annibale and Lohse have recently suggested a way1 in which the two-dimensional fluorescence intensity fluctuation (2D FIF) spectrometry2 may be further refined. Their main suggestion is to include a step in the analysis process where a case-by-case inspection of individual regions of interest of a membrane allows for selection of portions of the membrane which are "as homogenous as possible" and thereby exclude intensity spots potentially related to other sub-cellular structures. By incorporating that proposal into an objective and reproducible algorithm, here we show that 2D FIF has a built-in capability to automatically filter out such contributions, and that further removal of inhomogeneities does not alter the final results.

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