Combining 3D Magnetic Force Actuator and Multi-Functional Fluorescence Imaging to Study Nucleus Mechanobiology

作者： Miao Huang1,5, Heyang Wang1,5, Alfredo A. Delgado1, Tyler A. Reid1, Julian Long2, Shu Wang3,5, Hayley Sussman4, Juan Guan5,6,7, Hitomi Yamaguchi1, Xin Tang1,5,8,9 1Department of Mechanical and Aerospace Engineering, Herbert Wertheim College of Engineering,University of Florida, 2Department of Materials Science and Engineering,University of Florida, 3Department of Biostatistics,University of Florida, 4Department of Biomedical Engineering, College of Engineering (COE),University of Delaware (UD), 5UF Health Cancer Center,University of Florida, 6Department of Physics, College of Liberal Arts and Sciences,University of Florida, 7Department of Anatomy and Cell Biology, College of Medicine,University of Florida, 8J. Crayton Pruitt Family Department of Biomedical Engineering,University of Florida, 9Department of Physiology and Functional Genomics,University of Florida

简介：

Overview

This study explores a new protocol for applying mechanical force directly to the cell nucleus using magnetic microbeads within living cells, while enabling live-cell fluorescent imaging. This technique reveals insights into nuclear mechanosensing mechanisms, functioning without disrupting cellular processes.

Key Study Components

Research Area

• Cell biology
• Mechanobiology
• Imaging techniques

Background

• Understanding nuclear mechanosensing is crucial for cellular function.
• Existing techniques like optical tweezers are invasive and less efficient.
• Magnetic microbeads offer a non-invasive method to study these mechanisms.

Methods Used

• Application of mechanical force using magnetic microbeads.
• Live-cell imaging with confocal fluorescence microscopy.
• Quantitative imaging analysis to measure force-induced nuclear deformation.

Main Results

• Force application from microbeads led to nuclear deformation and YES-associated protein translocation.
• No significant differences in nuclear circularity between control cells and those with microbead internalization.
• The study confirms that magnetic force influences nuclear mechanotransduction pathways.

Conclusions

• This protocol advances the understanding of nuclear mechanosensing.
• It offers a non-invasive approach for investigating cellular mechanics and signaling pathways.

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