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. 2020 Mar 17;5(12):6947–6955. doi: 10.1021/acsomega.0c00322

Figure 2.

Figure 2

Fluorescent time traces of single Alexa 546–Alexa 647 FRET pairs with 13.6 nm (40 bp) separation diffusing in the confocal setup (a–c) and in a 110 nm-diameter ZMW (d–f) with a 0.5 ms binning time. The traces (a,d) show the donor emission after donor direct excitation at 557 nm, the traces (b,e) show the acceptor emission after acceptor direct excitation at 635 nm, and the traces (c,f) show the FRET emission (acceptor fluorescence) after donor excitation at 557 nm. The fluorescence enhancement in the ZMW directly leads to brighter detection events (d–f) as compared to the confocal reference (a–c) without any postprocessing.