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. 2016 Feb 10;7:10619. doi: 10.1038/ncomms10619

Figure 2. Single-molecule fluorescence characterization of enzyme encapsulation.

Figure 2

(a) Schematic illustration of single-molecule fluorescence co-localization of Cy3-labelled protein with Cy5-labelled cage using TIRF microscopy. DNA cages were captured on the surface by biotin-streptavidin interaction. (b) Representative field of view of enzyme-encapsulating cages under TIRF microscope. Examples of Cy3-Cy5 co-localization are highlighted using a pair of rectangles. Scale bar, 10 μm. (c) Quantified encapsulation yield for six different enzymes. The total number of molecules analysed for each protein is shown in Supplementary Table 2. The error bars represent the standard deviation obtained from the analysis of two to four movies of the sample from the same batch. (d) Fluorophore photobleaching trajectories with one, two, and three photobleaching steps. Photobleaching steps were quantitatively analysed by fitting the trajectories by HMM in QUB program. (e) Photobleaching statistics for Cy3-labelled proteins encapsulated within half-cages (Half[G6pDH]) or full-cages (Full[G6pDH]), as well as for an unencapsulated protein control (G6pDH). HMM, hidden-Markov modelling.