NanoTracker: Force-Sensing Optical Tweezers for Quantitative

In the past decade, experiments involving the manipulation and observation of nanostructures with light using optical tweezers methodology have developed from proof-of-principle experiments to an established quantitative technique in fields ranging from (bio)physics to cell biology. With optical tweezers, microscopically small objects can be held and manipulated. At the same time, the forces exerted on the trapped objects can be accurately measured. With the Prism-Award winning NanoTracker a platform for performing experiments using specimen from single molecules to whole cells is available. With two time-continuous traps, it allows the controlled trapping and accurate tracking of nanoparticles, suspended either in a microfluidic multichannel flow chamber or even in a temperature-controlled open Petri dish. With its 3D detection system, particle displacements in the trap can be recorded with nanometer precision. Moreover, dynamic forces acting on the particle can be measured with better than picoNewton resolution on a microsecond time-scale. Here, we discuss design features of and measurements done with the NanoTracker platform. In particular, we show how one of the hallmarks of single-molecule biophysics, the overstretching transition of DNA, can be studied in a versatile manner and used for protein-DNA interaction mechanics. Moreover, on the lower side of the force range the other benchmark single-molecule biophysics, kinesin’s 8-nm steps and stall forces, are shown to be measurable. With the NanoTracker, optical tweezers finally transcend from the labs of self-building scientists who helped the technique mature, to a turn-key system able to serve a much wider community of researchers in the life sciences