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Pulsed-Interleaved Excitation FRET Measurements on Single Duplex DNA Molecules Inside C-Shaped Nanoapertures

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journal contribution
posted on 03.04.2020, 14:58 by Samantha Fore, Yin Yuen, Lambertus Hesselink, Thomas Huser
Single-molecule fluorescence resonant energy transfer (FRET) is a widely accepted method for determining the spatial separation between molecules. In combination with pulsed interleaved excitation (PIE), additional information about the stoichiometry of molecular interactions is obtained. PIE-FRET, however, as implemented with standard confocal optics, requires the dilution of the sample to biologically low concentrations. Here, we show that PIE-FRET measurements inside nanometer-sized apertures yield meaningful biochemical data at 1000× higher concentrations.