Three-dimensional label-free observation of individual bacteria upon antibiotic treatment using optical diffraction tomography

AbstractMeasuring alterations in bacteria upon antibiotic application is important for basic studies in microbiology, drug discovery, and clinical diagnosis, and disease treatment. However, imaging and 3D time-lapse response analysis of individual bacteria upon antibiotic application remain largely unexplored mainly due to limitations in imaging techniques. Here, we present a method to systematically investigate the alterations in individual bacteria in 3D and quantitatively analyze the effects of antibiotics. Using optical diffraction tomography, in-situ responses of Escherichia coli and Bacillus subtilis to various concentrations of ampicillin were investigated in a label-free and quantitative manner. The presented method reconstructs the dynamic changes in the 3D refractive-index distributions of living bacteria in response to antibiotics at sub-micrometer spatial resolution.

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

Scientific Reports ◽

10.1038/s41598-018-20113-w ◽

2018 ◽

Vol 8 (1) ◽

Cited By ~ 12

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

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

Cells ◽

10.3390/cells8111368 ◽

2019 ◽

Vol 8 (11) ◽

pp. 1368 ◽

Cited By ~ 3

Author(s):

Kim ◽

Lee ◽

Fujii ◽

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.

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Three-Dimensional Label-Free Characterization of Frog Erythrocytes using Optical Diffraction Tomography

2018 IEEE Photonics Conference (IPC) ◽

10.1109/ipcon.2018.8527307 ◽

2018 ◽

Author(s):

Geon Kim ◽

Moosung Lee ◽

Daeheon Kwon ◽

SeongYeon Youn ◽

EuiTai Lee ◽

...

Keyword(s):

Three Dimensional ◽

Optical Diffraction ◽

Diffraction Tomography ◽

Label Free ◽

Optical Diffraction Tomography

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Detection of intracellular monosodium urate crystals in gout synovial fluid using optical diffraction tomography

Scientific Reports ◽

10.1038/s41598-021-89337-7 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Sangwoo Park ◽

Lucy Eunju Lee ◽

Hanna Kim ◽

Ji Eun Kim ◽

Seung Jun Lee ◽

...

Keyword(s):

Three Dimensional ◽

Time Lapse ◽

Optical Diffraction ◽

Diffraction Tomography ◽

Monosodium Urate ◽

Monosodium Urate Crystals ◽

Optical Diffraction Tomography ◽

Time Lapse Imaging ◽

Msu Crystals ◽

Urate Crystals

AbstractOptical diffraction tomography (ODT) enables imaging of unlabeled intracellular components by measuring the three-dimensional (3D) refractive index (RI). We aimed to detect intracellular monosodium urate (MSU) crystals in synovial leukocytes derived from gout patients using ODT. The 3D RI values of the synthetic MSU crystals, measured by ODT, ranged between 1.383 and 1.440. After adding synthetic MSU crystals to a macrophage, RI tomograms were reconstructed using ODT, and the reconstructed RI tomograms discerned intracellular and extracellular MSU crystals. We observed unlabeled synthetic MSU crystal entry into the cytoplasm of a macrophage through time-lapse imaging. Furthermore, using gout patient-derived synovial leukocytes, we successfully obtained RI tomogram images of intracellular MSU crystals. The 3D RI identification of MSU crystals was verified with birefringence through polarization-sensitive ODT measurements. Together, our results provide evidence that this novel ODT can identify birefringent MSU crystals in synovial leukocytes of patients with gout.

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Label-free three-dimensional observations and quantitative characterisation of on-chip vasculogenesis using optical diffraction tomography

10.1101/2020.01.01.892620 ◽

2020 ◽

Cited By ~ 1

Author(s):

Chungha Lee ◽

Seunggyu Kim ◽

Herve Hugonnet ◽

Moosung Lee ◽

Weisun Park ◽

...

Keyword(s):

Three Dimensional ◽

Optical Diffraction ◽

Diffraction Tomography ◽

Label Free ◽

Vascular Networks ◽

Optical Diffraction Tomography ◽

Quantitative Characterisation ◽

Vascular Structures ◽

On Chip ◽

Intrinsic Imaging

Label-free, three-dimensional (3D) quantitative observations of on-chip vasculogenesis were achieved using optical diffraction tomography. Exploiting 3D refractive index maps as an intrinsic imaging contrast, the vascular structures, multicellular activities, and subcellular organelles of endothelial cells were imaged and analysed throughout vasculogenesis to characterise mature vascular networks without exogenous labelling.

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Three-dimensional label-free observation of individual bacteria upon antibiotic treatment using optical diffraction tomography

Biomedical Optics Express ◽

10.1364/boe.377740 ◽

2020 ◽

Vol 11 (3) ◽

pp. 1257 ◽

Cited By ~ 2

Author(s):

Jeonghun Oh ◽

Jea Sung Ryu ◽

Moosung Lee ◽

Jaehwang Jung ◽

SeungYun Han ◽

...

Keyword(s):

Antibiotic Treatment ◽

Three Dimensional ◽

Optical Diffraction ◽

Diffraction Tomography ◽

Label Free ◽

Optical Diffraction Tomography

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

10.1101/219378 ◽

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.

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