Volumetric label-free histopathology using optical diffraction tomography

2021 ◽  
Author(s):  
Herve Hugonnet ◽  
Ariel J. Lee ◽  
Yeon Wook Kim ◽  
Moosung Lee ◽  
Seungwoo Shin ◽  
...  
Keyword(s):  
Optical Diffraction ◽  
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 Label-free monitoring of 3D cortical neuronal growth in vitro using optical diffraction tomography

2021 ◽  
Author(s):  
Ariel J. Lee ◽  
DongJo Yoon ◽  
SeungYun Han ◽  
Herve Hugonnet ◽  
WeiSun Park ◽  
...  
Keyword(s):  
Optical Diffraction ◽  
Neuronal Cultures ◽  
Multiple Time ◽  
Diffraction Tomography ◽  
Label Free ◽  
Neuronal Growth ◽  
Monitoring Method ◽  
Optical Diffraction Tomography ◽  
Immunofluorescence Labeling

The highly complex central nervous systems of mammals are often studied using three-dimensional (3D) in vitro primary neuronal cultures. A coupled confocal microscopy and immunofluorescence labeling are widely utilized for visualizing the 3D structures of neurons. However, this requires fixation of the neurons and is not suitable for monitoring an identical sample at multiple time points. Thus, we propose a label-free monitoring method for 3D neuronal growth based on refractive index tomograms obtained by optical diffraction tomography. The 3D morphology of the neurons was clearly visualized, and the developmental processes of neurite outgrowth in 3D spaces were analyzed for individual neurons.

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

2019 ◽  
Author(s):  
Jeonghun Oh ◽  
Jea Sung Ryu ◽  
Moosung Lee ◽  
Jaehwang Jung ◽  
Seung yun Han ◽  
...  
Keyword(s):  
Imaging Techniques ◽  
Three Dimensional ◽  
Time Lapse ◽  
Optical Diffraction ◽  
Response Analysis ◽  
Diffraction Tomography ◽  
Label Free ◽  
Optical Diffraction Tomography ◽  
Quantitative Manner ◽  
Basic Studies

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.

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

2020 ◽  
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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