Advances in design and testing of limited angle optical diffraction tomography system for biological applications

Abstract

Optical diffraction tomography has been steadily proving its potential to study one of the hot topics in modern cell biology — 3D dynamic changes in cells' morphology represented with refractive index values. In this technique digital holography is combined with tomographic reconstruction and thus it is necessary to provide projections acquired at different viewing directions. Usually the Mach-Zehnder interferometer configuration is used and while the object beam performs scanning, the reference beam is in most cases stationary. This approach either limits possible scanning strategies or requires additional mechanical movement to be introduced in the reference beam. On the other hand, spiral or grid scanning is possible in alternative common-path or Michelson configurations. However, in this case there is no guarantee that a specimen is sparse enough for the object to interfere with an object-free part of the beam. In this paper we present a modified version of Mach-Zehnder interferometer-based tomographic microscope, in which both object and reference beam are subject to scanning using one scanning device only thus making any scanning scenario possible. This concept is realized with a custom-built optical system in the reference beam and is appropriate for mechanical as well as optical scanning. Usually, the tomographic reconstruction setups and algorithms are verified using a microsphere phantom, which is not enough to test the influence of the distribution of the projections. In this work we propose a more complex calibration object created using two-photon polymerization.