Lab-on-Chip for fast 3D particle tracking in living cells

Abstract

We present a novel Lab-on-Chip technology for 3D particle tracking in living cells based on V-shaped micro-mirrors, which are used to observe fluorescent specimens from multiple vantage points, providing simultaneous stereo-images that can be recombined for 3D reconstruction. Our technology can be readily used with standard fluorescence microscopes, and we apply it to study chromatin dynamics using yeast strains with one or two GFP-tagged gene loci. Using an Andor EMCCD camera, loci are followed in 3D with inter-frame intervals of up to 10 ms and with an error of 27 nm per axis, yielding quantitative information on their dynamics with exquisite temporal spatial resolution.