Dual-wavelength cell’s refractive index analysis with partially coherent optical diffraction tomography

Author(s):  
J. M. Soto ◽  
J. A. Rodrigo ◽  
T. Alieva
2021 ◽  
Vol 9 ◽  
Author(s):  
Juan M. Soto ◽  
José A. Rodrigo ◽  
Tatiana Alieva

Optical diffraction tomography (ODT) is a computational imaging technique based on refractive index (RI) contrast. Its application for microscopic imaging of weakly absorbing and scattering samples has been demonstrated by using a specially designed holographic microscope with angular scanning of the coherent sample illumination direction. Recently, an alternative low cost technique based on partially coherent sample illumination (PC-ODT), which is compatible with the conventional wide-field transmission microscope, has been established. In this case, the 3D refractive index distribution of the sample is obtained by deconvolution from a single stack of through-focus intensity images. The performance of PC-ODT has been successfully tested on various fixed specimens (diatom frustule and biological cells) and moving bacteria. Here, we demonstrate that the PC-ODT is an efficient tool for the analysis of living eukaryotic cell dynamics at short- and long-term periods. The COS-7 cells, which hail from the African green monkey kidney, have been chosen for this study. A fast data acquisition setup comprising an optical scanning module can be easily attached to the microscope, and it allows observing cell 3D organelle movements and RI variations, with the required temporal resolution. In particular, a more rapid nucleoli rotation than previously reported has been found. The long-term cell monitoring during necrosis reveals significant changes in cell dry mass concentration obtained from recovered RI contrast.


Lab on a Chip ◽  
2018 ◽  
Vol 18 (22) ◽  
pp. 3484-3491 ◽  
Author(s):  
Seungwoo Shin ◽  
Jihye Kim ◽  
Je-Ryung Lee ◽  
Eun-chae Jeon ◽  
Tae-Jin Je ◽  
...  

Resolution-enhanced optical diffraction tomography using a micromirror-embedded coverslips.


2018 ◽  
Author(s):  
Chansuk Park ◽  
SangYun Lee ◽  
Geon Kim ◽  
SeungJun Lee ◽  
Jaehoon Lee ◽  
...  

Three-dimensional (3D) refractive index (RI) imaging and quantitative analyses of angiosperm pollen grains are presented. Using optical diffraction tomography, the 3D RI structures of individual angiosperm pollen grains were measured without using labeling or other preparation techniques. Various physical quantities, including volume, surface area, exine volume, and sphericity, were determined from the measured RI tomograms of pollen grains. Exine skeletons, the distinct internal structures of angiosperm pollen grains, were identified and systematically analyzed.


2018 ◽  
Vol 36 (24) ◽  
pp. 5754-5763 ◽  
Author(s):  
Shengli Fan ◽  
Seth Smith-Dryden ◽  
Jian Zhao ◽  
Stefan Gausmann ◽  
Axel Schulzgen ◽  
...  

2007 ◽  
Vol 15 (2) ◽  
Author(s):  
T. Kozacki ◽  
M. Kujawińska ◽  
P. Kniażewski

AbstractOptical diffraction tomography (ODT) applied to measurement of optical microelements is limited by low dynamic range, i.e., only objects with small deviations of refractive-index distribution can be measured. Therefore in this paper the limitations and errors of ODT are investigated throughout extensive numerical experiments. It is shown that these errors can be reduced by introduction of additional numerical focusing in the tomographic reconstruction algorithm. Additionally, new tomographic reconstruction algorithm using back propagation in reference medium for optical microelements measurement with known design is proposed. This hybrid reconstruction algorithm allows significant extension of ODT applicability in measurement of elements having large deviations of refractive-index distribution.


2017 ◽  
Author(s):  
Geon Kim ◽  
SangYun Lee ◽  
Seungwoo Shin ◽  
YongKeun Park

SummaryThe structure of pollen grains is related to the reproductive function of the plants. Here, three-dimensional (3D) refractive index maps were obtained for individual conifer pollen grains using optical diffraction tomography (ODT).The 3D morphological features of pollen grains from pine trees were investigated using measured refractive index maps, in which distinct substructures were clearly distinguished and analyzed.Morphological and physiochemical parameters of the pollen grains were quantified from the obtained refractive index (RI) maps and used to quantitatively study the interspecific differences of pollen grains from different strains.Our results demonstrate that ODT can assess the structure of pollen grains. This label-free and rapid 3D imaging approach may provide a new platform for understanding the physiology of pollen grains.


2016 ◽  
Author(s):  
Doyeon Kim ◽  
Nuri Oh ◽  
Kyoohyun Kim ◽  
SangYun Lee ◽  
Chan-Gi Pack ◽  
...  

AbstractDelivery of gold nanoparticles (GNPs) into live cells has high potentials, ranging from molecular-specific imaging, photodiagnostics, to photothermal therapy. However, studying the long-term dynamics of cells with GNPs using conventional fluorescence techniques suffers from phototoxicity and photobleaching. Here, we present a method for 3-D imaging of GNPs inside live cells exploiting refractive index (RI) as imaging contrast. Employing optical diffraction tomography, 3-D RI tomograms of live cells with GNPs are precisely measured for an extended period with sub-micrometer resolution. The locations and contents of GNPs in live cells are precisely addressed and quantified due to their distinctly high RI values, which was validated by confocal fluorescence imaging of fluorescent dye conjugated GNPs. In addition, we perform quantitative imaging analysis including the segmentations of GNPs in the cytosol, the volume distributions of aggregated GNPs, and the temporal evolution of GNPs contents in HeLa and 4T1 cells.AbbreviationsGNPsgold nanoparticlesRIrefractive indexODToptical diffraction tomographyDMDdigital micromirror device


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