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Issue 4, 2021

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

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Abstract

Biophysical properties of cells such as intracellular mass density and cell mechanics are known to be involved in a wide range of homeostatic functions and pathological alterations. An optical readout that can be used to quantify such properties is the refractive index (RI) distribution. It has been recently reported that the nucleus, initially presumed to be the organelle with the highest dry mass density (ρ) within the cell, has in fact a lower RI and ρ than its surrounding cytoplasm. These studies have either been conducted in suspended cells, or cells adhered on 2D substrates, neither of which reflects the situation in vivo where cells are surrounded by the extracellular matrix (ECM). To better approximate the 3D situation, we encapsulated cells in 3D covalently-crosslinked alginate hydrogels with varying stiffness, and imaged the 3D RI distribution of cells, using a combined optical diffraction tomography (ODT)-epifluorescence microscope. Unexpectedly, the nuclei of cells in 3D displayed a higher ρ than the cytoplasm, in contrast to 2D cultures. Using a Brillouin-epifluorescence microscope we subsequently showed that in addition to higher ρ, the nuclei also had a higher longitudinal modulus (M) and viscosity (η) compared to the cytoplasm. Furthermore, increasing the stiffness of the hydrogel resulted in higher M for both the nuclei and cytoplasm of cells in stiff 3D alginate compared to cells in compliant 3D alginate. The ability to quantify intracellular biophysical properties with non-invasive techniques will improve our understanding of biological processes such as dormancy, apoptosis, cell growth or stem cell differentiation.

Graphical abstract: Optical quantification of intracellular mass density and cell mechanics in 3D mechanical confinement

Supplementary files

Article information


Submitted
28 Aug 2020
Accepted
16 Nov 2020
First published
20 Nov 2020

This article is Open Access

Soft Matter, 2021,17, 853-862
Article type
Paper

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

S. Bakhshandeh, H. M. Taïeb, R. Schlüßler, K. Kim, T. Beck, A. Taubenberger, J. Guck and A. Cipitria, Soft Matter, 2021, 17, 853 DOI: 10.1039/D0SM01556C

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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