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1. Modeling stem cell nucleus mechanics using confocal microscopy

   https://doi.org/10.1007/s10237-021-01513-w  

   Kennedy, Zeke; Newberg, Joshua; Goelzer, Matthew; Judex, Stefan; Fitzpatrick, Clare K.; Uzer, Gunes (August 2021, Biomechanics and Modeling in Mechanobiology)

   null (Ed.)

   Nuclear mechanics is emerging as a key component of stem cell function and differentiation. While changes in nuclear structure can be visually imaged with confocal microscopy, mechanical characterization of the nucleus and its sub-cellular components require specialized testing equipment. A computational model permitting cell-specific mechanical information directly from confocal and atomic force microscopy of cell nuclei would be of great value. Here, we developed a computational framework for generating finite element models of isolated cell nuclei from multiple confocal microscopy scans and simple atomic force microscopy (AFM) tests. Confocal imaging stacks of isolated mesenchymal stem cells were converted into finite element models and siRNA-mediated Lamin A/C depletion isolated chromatin and Lamin A/C structures. Using AFM-measured experimental stiffness values, a set of conversion factors were determined for both chromatin and Lamin A/C to map the voxel intensity of the original images to the element stiffness, allowing the prediction of nuclear stiffness in an additional set of other nuclei. The developed computational framework will identify the contribution of a multitude of sub-nuclear structures and predict global nuclear stiffness of multiple nuclei based on simple nuclear isolation protocols, confocal images and AFM tests. 

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2. Impact of Silk-Ionomer Encapsulation on Immune Cell Mechanical Properties and Viability

   https://doi.org/10.1021/acsbiomaterials.4c00412  

   Kumarasinghe, Udathari; Hasturk, Onur; Wang, Brook; Rudolph, Sara; Chen, Ying; Kaplan, David L; Staii, Cristian (July 2024, ACS Biomaterials Science & Engineering)

   Encapsulation of single cells is a powerful technique used in various fields, such as regenerative medicine, drug delivery, tissue regeneration, cell-based therapies, and biotechnology. It offers a method to protect cells by providing cytocompatible coatings to strengthen cells against mechanical and environmental perturbations. Silk fibroin, derived from the silkworm Bombyx mori, is a promising protein biomaterial for cell encapsulation due to the cytocompatibility and capacity to maintain cell functionality. Here, THP-1 cells, a human leukemia monocytic cell line, were encapsulated with chemically modified silk polyelectrolytes through electrostatic layer-by-layer deposition. The effectiveness of the silk nanocoating was assessed using scanning electron microscopy (SEM) and confocal microscopy and on cell viability and proliferation by Alamar Blue assay and live/dead staining. An analysis of the mechanical properties of the encapsulated cells was conducted using atomic force microscopy (AFM) nanoindentation to measure elasticity maps and cellular stiffness. After the cells were encapsulated in silk, an increase in their stiffness was observed. Based on this observation, we developed a mechanical predictive model to estimate the variations in stiffness in relation to the thickness of the coating. By tuning the cellular assembly and biomechanics, these encapsulations promote systems that protect cells during biomaterial deposition or processing in general. 

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3. Brillouin microscopy monitors rapid responses in subcellular compartments

   https://doi.org/10.1186/s43074-024-00123-w  

   Coker, Zachary N; Troyanova-Wood, Maria; Steelman, Zachary A; Ibey, Bennett L; Bixler, Joel N; Scully, Marlan O; Yakovlev, Vladislav V (December 2024, PhotoniX)

   Abstract Measurements and imaging of the mechanical response of biological cells are critical for understanding the mechanisms of many diseases, and for fundamental studies of energy, signal and force transduction. The recent emergence of Brillouin microscopy as a powerful non-contact, label-free way to non-invasively and non-destructively assess local viscoelastic properties provides an opportunity to expand the scope of biomechanical research to the sub-cellular level. Brillouin spectroscopy has recently been validated through static measurements of cell viscoelastic properties, however, fast (sub-second) measurements of sub-cellular cytomechanical changes have yet to be reported. In this report, we utilize a custom multimodal spectroscopy system to monitor for the very first time the rapid viscoelastic response of cells and subcellular structures to a short-duration electrical impulse. The cytomechanical response of three subcellular structures - cytoplasm, nucleoplasm, and nucleoli - were monitored, showing distinct mechanical changes despite an identical stimulus. Through this pioneering transformative study, we demonstrate the capability of Brillouin spectroscopy to measure rapid, real-time biomechanical changes within distinct subcellular compartments. Our results support the promising future of Brillouin spectroscopy within the broad scope of cellular biomechanics. 

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4. Measuring viscoelasticity of soft biological samples using atomic force microscopy

   https://doi.org/10.1039/c9sm01020c  

   Efremov, Yuri M.; Okajima, Takaharu; Raman, Arvind (January 2020, Soft Matter)

   Mechanical properties play important roles at different scales in biology. At the level of a single cell, the mechanical properties mediate mechanosensing and mechanotransduction, while at the tissue and organ levels, changes in mechanical properties are closely connected to disease and physiological processes. Over the past three decades, atomic force microscopy (AFM) has become one of the most widely used tools in the mechanical characterization of soft samples, ranging from molecules, cell organoids and cells to whole tissue. AFM methods can be used to quantify both elastic and viscoelastic properties, and significant recent developments in the latter have been enabled by the introduction of new techniques and models for data analysis. Here, we review AFM techniques developed in recent years for examining the viscoelastic properties of cells and soft gels, describe the main steps in typical data acquisition and analysis protocols, and discuss relevant viscoelastic models and how these have been used to characterize the specific features of cellular and other biological samples. We also discuss recent trends and potential directions for this field. 

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5. Mechanical spectroscopy of materials using atomic force microscopy (AFM-MS)

   https://doi.org/10.1016/j.mattod.2024.08.021  

   Petrov, M; Canena, D; Kulachenkov, N; Kumar, N; Nickmilder, Pierre; Leclère, Philippe; Sokolov, Igor (November 2024, Materials Today)

   Here, we present a novel mechano-spectroscopic atomic force microscopy (AFM-MS) technique that overcomes the limitations of current spectroscopic methods by combining the high-resolution imaging capabilities of AFM with machine learning (ML) classification. AFM-MS employs AFM operating in sub-resonance tapping imaging mode, which enables the collection of multiple physical and mechanical property maps of a sample with sub-nanometer lateral resolution in a highly repeatable manner. By comparing these properties to a database of known materials, the technique identifies the location of constituent materials at each image pixel with the assistance of ML algorithms. We demonstrate AFM-MS on various material mixtures, achieving an unprecedented lateral spectroscopic resolution of 1.6 nm. This powerful approach opens new avenues for nanoscale material study, including the material identification and correlation of nanostructure with macroscopic material properties. The ability to map material composition with such high resolution will significantly advance the understanding and design of complex, nanostructured materials. 

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