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Published on: November 2, 2018
In Situ Quantitative Imaging of Plasma Membrane Stiffness in Live Cells Using a Genetically Encoded FRET Sensor
Yusi Hu1, Hai-Yan Wen2, Meng-Yao Liu1
1State Key Laboratory of Medicinal Chemical Biology, Frontiers Science Center for Cell Responses, Tianjin Key Laboratory of Biosensing and Molecular Recognition, Research Centre for Analytical Sciences, College of Chemistry and School of Medicine, Nankai University, Tianjin 300071, P. R. China.
We developed a novel FRET sensor to measure cell membrane stiffness, crucial for cellular function. This tool tracks cholesterol and sphingomyelin proximity, offering insights into mechanobiology and viral interactions.
Area of Science:
- Cellular Biology
- Biophysics
- Molecular Biology
Background:
- Cell membrane stiffness is vital for cellular processes.
- Cholesterol and sphingomyelin are key regulators of membrane rigidity.
- Existing methods for measuring membrane stiffness are limited by invasiveness and speed.
Purpose of the Study:
- To develop and validate a novel, non-invasive FRET-based sensor for measuring cell membrane stiffness.
- To investigate the impact of various physiological conditions on plasma membrane stiffness.
- To explore the effects of SARS-CoV-2 infection on cell membrane properties.
Main Methods:
- Development of a FRET-based protein sensor targeting cholesterol and sphingomyelin proximity.
- Utilizing confocal microscopy for real-time FRET signal detection.
- Applying the sensor to assess membrane stiffness under osmotic stress, reactive oxygen species (ROS) exposure, and varying substrate stiffness.
Main Results:
- The FRET sensor effectively reports on membrane stiffness by detecting cholesterol-sphingomyelin proximity.
- Plasma membrane stiffness was shown to change under different osmotic pressures, ROS levels, and substrate stiffness.
- The study observed alterations in membrane stiffness and ACE2 distribution upon SARS-CoV-2 attachment.
Conclusions:
- The developed FRET sensor provides a sensitive and adaptable tool for studying cell membrane mechanics.
- This technology enables real-time assessment of membrane stiffness in response to diverse stimuli.
- The findings highlight the potential of this sensor for investigating viral-host interactions and mechanobiology.
