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Long-Time Plasma Membrane Imaging Based on a Two-Step Synergistic Cell Surface Modification Strategy.

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  • 1State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University , Nanjing 210096, P. R. China.

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Summary

We developed a novel two-step cell surface modification strategy for long-time plasma membrane imaging. This method overcomes dye internalization and detachment issues, enabling stable imaging for up to 8 hours.

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Materials Science

Background:

  • Long-time plasma membrane imaging is challenging due to rapid dye internalization and detachment.
  • Existing imaging reagents like DiD and CellMask have limitations in stability and duration.

Purpose of the Study:

  • To develop a novel, stable, and long-lasting plasma membrane imaging strategy.
  • To overcome the limitations of current plasma membrane imaging techniques.

Main Methods:

  • A two-step synergistic cell surface modification and labeling strategy was employed.
  • Cells were treated with glycol chitosan-10% PEG2000 cholesterol-10% biotin (GC-Chol-Biotin) for membrane anchoring.
  • Fluorescein isothiocyanate (FITC)-conjugated avidin was used for fluorescence labeling via biotin-avidin recognition.

Main Results:

  • Achieved stable plasma membrane imaging for up to 8 hours without significant dye internalization or detachment.
  • Demonstrated superior imaging performance compared to commercial reagents DiD and CellMask.
  • Successfully tracked photodynamic damage to plasma membranes (caused by Chlorin e6) in real-time for 5 hours.

Conclusions:

  • The developed imaging strategy provides a stable and long-lasting platform for plasma membrane observation.
  • This method enables dynamic recording of plasma membrane behaviors, including cell shrinkage, blebbing, and vesiculation.
  • Offers a novel approach for investigating plasma membrane dynamics over extended periods.