Fig. 1: Membrane topography variations can cause the appearance of clustering. | Communications Biology

Fig. 1: Membrane topography variations can cause the appearance of clustering.

From: Membrane topography and the overestimation of protein clustering in single molecule localisation microscopy – identification and correction

Fig. 1

a Membrane cross section of a simulated SMLM dataset of a membrane protein and a randomly distributed membrane marker, 716 × 193 pixels. b The dataset after smoothing with a Gaussian filter (sigma 24 pixels). The images were normalised to the same mean intensity and displayed with the same intensity range. In the protein image three clusters are apparent and the least prominent and its corresponding area in the membrane image marked (white circles). The corresponding areas in the datasets in a are marked with red circles. c The difference image [protein-membrane marker] between the two normalized Gaussian filtered images. The middle and originally least prominent of the three clusters in b (white circle) is now seen to be genuine, whereas the two larger clusters coincided with areas rich in membrane. Note, the display has been stretched to use the full range of the look up table where zero is at the centre. d The rendered membrane with folds used for the simulations.

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