SUPLEMENTAL DATA 1 (S1) : Addendum to Experimental Methods
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A 488 nm argon laser (Melles Griot) was used as the excitation source and the laser beam was focused using a 100X, 1.3 N.A., oil immersion apochromatic objective (Olympus). Throughout experiments excitation laser power was kept at 150 μW using neutral density filters. The emitted fluorescence was then collected by the same objective and an initial dichroic mirror (505DCXE, Chroma Technology) selected wavelengths > 505 nm. A 50 μm pinhole (Melles Griot) rejected out of focus fluorescence and a second dichroic mirror (D625LP, Chroma Technology) separated donor (QD605) and acceptor (Cy5) emission. Two avalanche photodiodes (APD, EG&G) detected signal from the two channels, which were filtered with separate bandpass filters (D605/20 and 660DF50, Chroma Technology). Acquisition software, written in Labview, and a digital counter (National Instruments) were used to collect data from the APDs. 10
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