3-Dimensional tracking of particles poses a challenge in terms of accurately measuring the Z-axis position. Typically it requires the acquisition of multiple images in different focal planes (using expensive scanning devices) as a function of time. The Z-stacks thus obtained comprise the 4D dataset (x,y,z,t ) that the tracking information is usually extracted from. Acquisition of Z-stacks is time-consuming and reduces the time-resolution of the data.
The technique using the Dual View introduced by Paul Selvin’s group at the University of Illinois at Urbana Champaign simplifies the measurement considerably by employing simultaneous bifocal imaging with no moving parts.