scholarly journals Optical quantification of intracellular mass density and cell mechanics in 3D mechanical confinement

Soft Matter ◽  
2021 ◽  
Author(s):  
Sadra Bakhshandeh ◽  
Hubert M. Taïeb ◽  
Raimund Schlüßler ◽  
Kyoohyun Kim ◽  
Timon Beck ◽  
...  

Optical quantification of intracellular mass density using optical diffraction tomography (ODT) and cell mechanics using Brillouin spectroscopy under 3D mechanical confinement.

2021 ◽  
Author(s):  
Kyoohyun Kim ◽  
Vamshidhar R. Gade ◽  
Teymuras V. Kurzchalia ◽  
Jochen Guck

Upon starvation or overcrowding, the nematode Caenorhabditis elegans enters diapause by forming a dauer larva. This larva can further transit into an anhydrobiotic state and survive harsh desiccation. We previously identified the genetic and biochemical pathways essential for survival — but without an accompanying physical model, the mechanistic understanding of this amazing phenomenon will remain inadequate. Neither microscopic investigation of structural changes upon entry into anhydrobiosis nor the most basic quantitative characterization of material properties of living desiccated larvae, however, have been feasible, due to lack of appropriate techniques. Here, we employed optical diffraction tomography (ODT) to quantitatively assess the internal mass density distribution of living larvae in the reproductive and diapause stages. More importantly, ODT allowed for the first time physical analysis of desiccated dauer larvae: their mass density was significantly increased in the anhydrobiotic state. We also applied ODT on different mutants that are sensitive to desiccation. Remarkably, one of them displayed structural abnormalities in the anhydrobiotic stage that could not be observed either by conventional light or electron microscopy. Our advance opens a door to quantitatively assessing fine differences in material properties and structure necessary to fully understanding an organism on the verge of life and death.


2021 ◽  
Author(s):  
Piotr Zdańkowski ◽  
Julianna Winnik ◽  
Paweł Gocłowski ◽  
Maciej Trusiak

Cells ◽  
2019 ◽  
Vol 8 (11) ◽  
pp. 1368 ◽  
Author(s):  
Kim ◽  
Lee ◽  
Fujii ◽  
Lee ◽  
Lee ◽  
...  

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.


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.


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