Monitoring the Brownian motion of individual nanoscopic objects is key to investigate their transport properties and interactions with their close environment. Most techniques rely on transient diffusion through a detection volume or immobilisation, which restrict observation times or motility. We measure the diffusion coefficient and surface charge of individual nanoparticles and DNA molecules in an anti-Brownian electrokinetic trap (ABELtrap). This instrument is an active feedback trap confining the Brownian motion of a nanoparticle to the detection site by applying an electric field based on the particle’s current position. We simulate the Brownian motion of nanospheres in our sample geometry, including wall effects, due to partial confinement in the third dimension. The theoretically predicted values are in excellent agreement with our diffusion measurements in the ABELtrap. We also demonstrate the ABELtrap’s ability to measure varying sizes of DNA origami structures during denaturation.
We have developed a workstation based on holographic tweezers to optically trap, move and characterize metal nanoparticles. Our advanced darkfield imaging system allows us to simultaneously image and take spectra of single trapped metal nanoparticles. We take advantage of the beamshaping abilities of the spatial light modulator and correct for aberrations of the trapping optics. We monitor the improvement of the optical trap with video-based nanoparticle tracking. Furthermore we theoretically assess the capabilities and limitations of video-based tracking for nanoparticle position detection, in particular with respect to acquisition frequencies below the corner frequency.
Combining smFRET and electrokinetic trapping to investigate dynamic conformational changes of single molecules in solution with millisecond time resolution for observation times extending beyond the duration of several reaction cycles.
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