Automatic three-dimensional tracking of particles with high-numerical-aperture digital lensless holographic microscopy

Abstract

We present an automatic procedure for 3D tracking of micrometer-sized particles with high-NA digital lensless holographic microscopy. The method uses a two-feature approach to search for the best focal planes and to distinguish particles from artifacts or other elements on the reconstructed stream of the holograms. A set of reconstructed images is axially projected onto a single image. From the projected image, the centers of mass of all the reconstructed elements are identified. Starting from the centers of mass, the morphology of the profile of the maximum intensity along the reconstruction direction allows for the distinguishing of particles from others elements. The method is tested with modeled holograms and applied to automatically track micrometer-sized bubbles in a sample of 4 mm3 of soda.

Review of digital holographic microscopy for three-dimensional profiling and tracking

“…Then, the axial position of the center of mass of each cell was determined from phase profiles at the focal plane. 101 Taking into account the morphological changes of cells, the concept of minimum boundary filters 100 was introduced to determine the positions of cells, and it was compared with other three methods, i.e., centroid, weighted centroid, and maximum phase value, as shown in Fig. 17.…”

Four-dimensional tracking of spatially incoherent illuminated samples using self-interference digital holography

“…Among them, 3D tracking of the sample in the entire field of view by DH is of great importance in diverse biologic and bio-technologic context [7]. Recently, many investigations focus on the ability of DH to track particles [8], cells [9] and clinical seminal samples [10] in the microfluidics channel. However, the reconstructed images suffer strong speckles in the highly coherent illuminated systems, as well as the relative high energy and focused laser beam maybe harmful to biological samples.…”

Holography: origin, basic principle and applications of a revolutionary communication method in art and science

“…It is also worth mentioning the interesting results regarding digital in-line holographic light microscopy developed to observe, with wavelength resolution, 3-D representations of the paths followed by micro objects such as suspensions of microspheres and biological samples (cells, bacteria, etc. ) and microchannel flow, motion of bubbles in microfluidic applications (Restrepo & García-Sucerquia, 2012).…”