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. 2014 May 12;9(5):e97440. doi: 10.1371/journal.pone.0097440

Figure 4. Segmented brightness analysis of monomers and dimers.

Figure 4

(A) Illustration of photodepletion for monomers and dimers. Fluorescent molecules are depicted as filled circles and photobleached molecules are pictured as broken circles. The normalized brightness of monomers (b = 1) remains unchanged by photodepletion, In contrast, photobleaching of a dimeric sample with initial brightness of 2 leads to a reduction of brightness as explained in the text. (B) Theoretical brightness based on segmentation analysis of a monomeric (solid line) and dimeric (dashed line) sample as a function of photodepletion. (C) Brightness of EGFP from a yeast cell by segmentation analysis versus photodepletion fraction. Brightness values (diamonds) for a data segment size of 1.6 s. Ten consecutive brightness values are averaged (red triangles) to better visualize the trend of the data. The dashed blue line represents a fit of the brightness values to Eq. 6 with a fitted value of Inline graphic. (D) Brightness of EGFP2 from a yeast cell by segmentation analysis versus photodepletion fraction. Symbols are described under (C). The blue dashed line represents a fit of the brightness values to Eq. 6 with a fitted value of Inline graphic.