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Updated: Jan 11, 2026
Live-cell Imaging of Migrating Cells Expressing Fluorescently-tagged Proteins in a Three-dimensional Matrix
Published on: December 22, 2011
A mechanosensitive fluorescent probe for visualizing migrasome mechano-spatiotemporal dynamics
Kongqi Chen1, Qiong Liu2, Mengjie Wang1
1School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong, China.
Abstract:
Migrasomes, recently identified as extracellular vesicles formed along the retraction fibers (RFs) in migrating cells, are evident in damaged mitochondria clearance, intercellular signaling, embryonic development, and pathological progression of diseases. During migrasome biogenesis, signaling cues and membrane dynamics interplay with each other in the maturation, expansion, and detachment of migrasomes from RFs. Membrane tension plays a crucial role in migrasome dynamics, and tools for monitoring the dynamic mechanical properties of migrasomes are highly desired. In this study, we developed and characterized a novel mechanosensitive but viscosity-insensitive fluorescent probe, named OT2SQ, for real-time investigation of migrasome dynamics in living cells. OT2SQ is a twisted small molecule with multiple rotatable units, exhibiting longer fluorescence lifetimes in response to increased membrane tension but showing no response to altered viscosity. Using time-lapse fluorescence lifetime imaging, we identified distinct modes of migrasome biogenesis and detachment from retraction fibers, each associated with unique membrane tension dynamics. Furthermore, phasor-FLIM analysis revealed alterations in migrasome membrane tension during content release and internalization processes. This study introduces a versatile fluorescent tool for real-time monitoring of migrasome morphology, spatiotemporal distribution, and membrane tension dynamics.
Insights
Researchers developed a novel fluorescent probe, OT2SQ, to measure membrane tension in migrasomes, crucial vesicles involved in cell repair and development. This tool enables real-time monitoring of migrasome dynamics and their mechanical properties.
Area of Science:
- Cell Biology
- Biophysics
- Molecular Imaging
Background:
- Migrasomes are extracellular vesicles involved in cellular processes like repair and development.
- Their biogenesis and function are influenced by membrane dynamics and tension.
- Existing tools lack the specificity to monitor migrasome membrane tension in real-time.
Purpose of the Study:
- To develop and characterize a novel fluorescent probe for real-time monitoring of migrasome membrane tension.
- To investigate the role of membrane tension in migrasome biogenesis, dynamics, and function.
Main Methods:
- Development of a novel mechanosensitive fluorescent probe (OT2SQ).
- Utilized time-lapse fluorescence lifetime imaging (FLIM) and phasor-FLIM analysis.
- Applied the probe to study migrasome dynamics in living cells.
Main Results:
- OT2SQ demonstrated viscosity-insensitivity and sensitivity to membrane tension.
- Distinct modes of migrasome biogenesis and detachment correlated with membrane tension changes.
- Alterations in migrasome membrane tension were observed during content release and internalization.
Conclusions:
- OT2SQ is a versatile tool for real-time monitoring of migrasome membrane tension.
- Provides insights into the mechanical properties governing migrasome dynamics.
- Facilitates further research into migrasome function in health and disease.
