KEYWORDS: Microfluidics, Refractive index, Tomography, Statistical analysis, Holography, Biological research, Lab on a chip, 3D metrology, Imaging systems
Holographic tomography (HT) is a label-free, high-resolution and non-invasive method that retrieves 3D refractive index (RI) information about analysed biological specimens. The most common measurement scenario includes culturing and analysing cells directly in a Petri dish. However, it does not mimic the in vivo conditions unlike the microfluidic approach. Thus, in our work, we have focused on the development of a measurement configuration that is dedicated to analysis of cell dynamics in a lab-on-chip. It includes a fast HT system, a new ultra-thin microfluidic chip that allows for long term monitoring in controlled environment, a stitching algorithm that allows to combine single fields of view (FoV) into a synthetic field of view in three dimensions and the full volume RI analysis of internal cellular organelles during measurements. This setup provides the ability to track changes occurring in individual cell organelles as well as getting statistically valuable data. In experimental verification, A549-type and MeWo cells were cultured under microfluidic conditions in the chip and put under observation using HT.
With increasing demand on data storage and transmission compression has became an important issue in the holographic measurement community. In this work we propose a lossy compression approach designed for the offaxis image plane holograms: a popular type of holograms in digital holographic microscopy and its derivatives. The method utilizes the band-limitation of the signal and is an extension for any conventional image codec. Several codecs are investigated and reliable compression parameters that maintain the holographic measurement precision are presented.
In this paper we discuss the influence of the camera noise in holographic projections measurements on the accuracy of reconstruction in the limited projection angle optical diffraction tomography (LAODT). To counteract the shortcomings of LAODT due to “missing cone” problem we apply generalized total variation iterative constraint (GTVIC) algorithm which replenishes the spectral contents of the reconstruction. To investigate the influence of the noise on result of the GTVIC reconstruction we perform systematic numerical experiments based on simulated phantom mimicking a cell and tailored to the measurement parameters of LAODT system. Next, to mitigate the disruptive influence of noise we test the efficiency of two denoising procedures based on blockmatching technique, namely BM3D and BM4D. Thanks to the properties of those algorithms, the denoising may be applied directly on holograms or hologram stacks, without destroying the fringes. The tomographic GTVIC reconstruction results from data after filtration will be compared with noise-free reconstruction, in reference to the simple median filtering of the noisy reconstruction.
In the paper the advantages of two different microscopic techniques, namely digital holographic microscopy (DHM) and Confocal Laser Scanning Microscopy (CLSM) have been combined with the aim to investigate HeLa cell culture in terms of statistical analysis of area of subcellular structures of HeLa cells and related to them dry mass estimation. To assure the proper statistical representation of the cells both measurements comprised of multiple fields of view (FoVs), stitched together to form two FoVs with overlapping regions. The results suggest a strong linear correlation of nucleoli dry mass to their projection area, a result that is promising in terms of its biological relevance.
Recovering phase information with Deterministic approaches as the Transport of Intensity Equation (TIE) has recently
emerged as an alternative tool to the interferometric techniques because it is experimentally easy to implement and
provides fast and accurate results. Moreover, the potential of employing partially coherent illumination (PCI) in such
techniques allow obtaining high quality phase reconstructions providing that the estimation of the corresponding Phase
Transfer Function (PTF) is carried out correctly. Hence, accurate estimation of the PTF requires that the physical
properties of the optical system are well known. Typically, these parameters are assumed constant in all the set of
measurements, which might not be optimal. In this work, we proposed the use of an amplitude Spatial Light Modulator
(aSLM) for tuning the degree of coherence of the optical system. The aSLM will be placed at the Fourier plane of the
optical system, and then, band pass filters will be displayed. This methodology will perform amplitude modulation of the
propagated field and as a result, the state of coherence of the optical system can be modified. Theoretical and
experimental results that validate our proposed technique will be shown.
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