scholarly journals Label-free high-resolution 3-D imaging of gold nanoparticles inside live cells using optical diffraction tomography

2016 ◽  
Author(s):  
Doyeon Kim ◽  
Nuri Oh ◽  
Kyoohyun Kim ◽  
SangYun Lee ◽  
Chan-Gi Pack ◽  
...  
Keyword(s):  
Gold Nanoparticles ◽  
Refractive Index ◽  
Quantitative Imaging ◽  
Extended Period ◽  
Optical Diffraction ◽  
Live Cells ◽  
Diffraction Tomography ◽  
Label Free ◽  
Optical Diffraction Tomography ◽  
4T1 Cells

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

scholarly journals Label-free high-resolution 3-D imaging of gold nanoparticles inside live cells using optical diffraction tomography

Methods ◽  
2018 ◽  
Vol 136 ◽  
pp. 160-167 ◽  
Author(s):  
Doyeon Kim ◽  
Nuri Oh ◽  
Kyoohyun Kim ◽  
SangYun Lee ◽  
Chan-Gi Pack ◽  
...  
Keyword(s):  
Gold Nanoparticles ◽  
High Resolution ◽  
Optical Diffraction ◽  
Live Cells ◽  
Diffraction Tomography ◽  
Label Free ◽  
Optical Diffraction Tomography

scholarly journals Mitotic Chromosomes in Live Cells Characterized Using High-Speed and Label-Free Optical Diffraction Tomography

Cells ◽  
2019 ◽  
Vol 8 (11) ◽  
pp. 1368 ◽  
Author(s):  
Kim ◽  
Lee ◽  
Fujii ◽  
Lee ◽  
Lee ◽  
...  
Keyword(s):  
Diffusion Coefficient ◽  
Three Dimensional ◽  
Laser Scanning Microscopy ◽  
Optical Diffraction ◽  
Live Cells ◽  
Confocal Laser ◽  
Diffraction Tomography ◽  
Label Free ◽  
Mitotic Chromosomes ◽  
Optical Diffraction Tomography

The cell nucleus is a three-dimensional, dynamic organelle organized into subnuclear compartments such as chromatin and nucleoli. The structure and function of these compartments are maintained by diffusion and interactions between related factors as well as by dynamic and structural changes. Recent studies using fluorescent microscopic techniques suggest that protein factors can access and are freely mobile in heterochromatin and in mitotic chromosomes, despite their densely packed structure. However, the physicochemical properties of the chromosome during cell division are not fully understood. In the present study, characteristic properties such as the refractive index (RI), volume of the mitotic chromosomes, and diffusion coefficient (D) of fluorescent probes inside the chromosome were quantified using an approach combining label-free optical diffraction tomography with complementary confocal laser-scanning microscopy and fluorescence correlation spectroscopy. Variations in these parameters correlated with osmotic conditions, suggesting that changes in RI are consistent with those of the diffusion coefficient for mitotic chromosomes and cytosol. Serial RI tomography images of chromosomes in live cells during mitosis were compared with three-dimensional confocal micrographs to demonstrate that compaction and decompaction of chromosomes induced by osmotic change were characterized by linked changes in chromosome RI, volume, and the mobilities of fluorescent proteins.

scholarly journals Correlative three-dimensional fluorescence and refractive index tomography: bridging the gap between molecular specificity and quantitative bioimaging

2017 ◽  
Author(s):  
Kyoohyun Kim ◽  
Wei Sun Park ◽  
Sangchan Na ◽  
Sangbum Kim ◽  
Taehong Kim ◽  
...  
Keyword(s):  
Refractive Index ◽  
Three Dimensional ◽  
Optical Diffraction ◽  
Live Cells ◽  
Specific Information ◽  
Label Free ◽  
Optical Diffraction Tomography ◽  
Molecular Specificity ◽  
3D Fluorescence Microscopy ◽  
Nih 3T3

AbstractOptical diffraction tomography (ODT) provides label-free three-dimensional (3D) refractive index (RI) measurement of biological samples. However, due to the nature of the RI values of biological specimens, ODT has limited access to molecular specific information. Here, we present an optical setup combining ODT with three-channel 3D fluorescence microscopy, to enhance the molecular specificity of the 3D RI measurement. The 3D RI distribution and 3D deconvoluted fluorescence images of HeLa cells and NIH-3T3 cells are measured, and the cross-correlative analysis between RI and fluorescence of live cells are presented.

scholarly journals 3D label-free imaging and analysis of Pinus pollen grains using optical diffraction tomography

2017 ◽  
Author(s):  
Geon Kim ◽  
SangYun Lee ◽  
Seungwoo Shin ◽  
YongKeun Park
Keyword(s):  
Refractive Index ◽  
Three Dimensional ◽  
Pollen Grains ◽  
Reproductive Function ◽  
Optical Diffraction ◽  
List Type ◽  
Diffraction Tomography ◽  
Label Free ◽  
Optical Diffraction Tomography ◽  
Index Maps

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.

scholarly journals Intracellular Detection and Localization of Nanoparticles by Refractive Index Measurement

Sensors ◽  
2021 ◽  
Vol 21 (15) ◽  
pp. 5001
Author(s):  
Alain Géloën ◽  
Karyna Isaieva ◽  
Mykola Isaiev ◽  
Olga Levinson ◽  
Emmanuelle Berger ◽  
...  
Keyword(s):  
Refractive Index ◽  
Optical Diffraction ◽  
Live Cells ◽  
Diffraction Tomography ◽  
Refractive Index Measurement ◽  
Optical Diffraction Tomography ◽  
Fluorescence Measurements ◽  
Index Measurement ◽  
Intracellular Detection

The measuring of nanoparticle toxicity faces an important limitation since it is based on metrics exposure, the concentration at which cells are exposed instead the true concentration inside the cells. In vitro studies of nanomaterials would benefit from the direct measuring of the true intracellular dose of nanoparticles. The objective of the present study was to state whether the intracellular detection of nanodiamonds is possible by measuring the refractive index. Based on optical diffraction tomography of treated live cells, the results show that unlabeled nanoparticles can be detected and localized inside cells. The results were confirmed by fluorescence measurements. Optical diffraction tomography paves the way to measuring the true intracellular concentrations and the localization of nanoparticles which will improve the dose-response paradigm of pharmacology and toxicology in the field of nanomaterials.

scholarly journals Three-dimensional label-free imaging and analysis of Pinus pollen grains using optical diffraction tomography

2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Geon Kim ◽  
SangYun Lee ◽  
Seungwoo Shin ◽  
YongKeun Park
Keyword(s):  
Three Dimensional ◽  
Pollen Grains ◽  
Optical Diffraction ◽  
Diffraction Tomography ◽  
Label Free ◽  
Optical Diffraction Tomography

Enhancement of optical resolution in three-dimensional refractive-index tomograms of biological samples by employing micromirror-embedded coverslips

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 ◽  
...  
Keyword(s):  
Refractive Index ◽  
Biological Samples ◽  
Three Dimensional ◽  
Optical Resolution ◽  
Optical Diffraction ◽  
Diffraction Tomography ◽  
Optical Diffraction Tomography

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

Label-free analysis and identification of white blood cell population using optical diffraction tomography

2015 ◽  
Author(s):  
Jonghee Yoon ◽  
Kyoohyun Kim ◽  
Min-hyeok Kim ◽  
YoungJu Jo ◽  
Suk-Jo Kang ◽  
...  
Keyword(s):  
Blood Cell ◽  
Cell Population ◽  
White Blood Cell ◽  
Optical Diffraction ◽  
Diffraction Tomography ◽  
Label Free ◽  
Optical Diffraction Tomography

scholarly journals Three-dimensional refractive index distributions of individual angiosperm pollen grains

2018 ◽  
Author(s):  
Chansuk Park ◽  
SangYun Lee ◽  
Geon Kim ◽  
SeungJun Lee ◽  
Jaehoon Lee ◽  
...  
Keyword(s):  
Refractive Index ◽  
Three Dimensional ◽  
Pollen Grains ◽  
Optical Diffraction ◽  
Diffraction Tomography ◽  
Optical Diffraction Tomography ◽  
Internal Structures ◽  
Quantitative Analyses ◽  
Angiosperm Pollen ◽  
Preparation Techniques

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.

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