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Related Experiment Videos

Monitoring cell movements and volume changes with pulse-mode scanning ion conductance microscopy.

P Happel1, G Hoffmann, S A Mann

  • 1Department of Molecular Neurobiochemistry, Ruhr-University Bochum, NC7-170, Universitätsstr 150, D-44780 Bochum, Germany.

Journal of Microscopy
|November 25, 2003
PubMed
Summary

This study introduces pulse-mode scanning ion conductance microscopy (SICM) for observing cell volume and membrane dynamics during locomotion. The technique allows for quantitative monitoring of cell shape changes over minutes to hours.

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Area of Science:

  • Cell Biology
  • Biophysics
  • Microscopy

Background:

  • Cell locomotion involves dynamic changes in cell volume and membrane shape.
  • Observing these dynamic processes requires high-resolution imaging techniques capable of long-term monitoring.

Purpose of the Study:

  • To develop and validate a pulse-mode scanning ion conductance microscopy (SICM) method for observing cell volume and membrane movements during cell locomotion.
  • To quantitatively monitor cell shape changes over extended periods (minutes to hours) with high spatial resolution.

Main Methods:

  • Utilized a modified pulse-mode SICM instrument with current pulses for precise electrode-to-surface distance control.
  • Implemented a back-step mode to prevent tip-membrane contact during lateral scanning.
  • Employed feedback-controlled piezoactuators for repeated cell surface scans.

Related Experiment Videos

  • Developed image subtraction software to quantify volume and positional changes between scans.
  • Applied line scan mode for high-temporal-resolution monitoring of specific cell sections.

    • Main Results:

    • Achieved reproducible height measurements of cultured cells with a standard deviation of 50 nm.
    • Quantitatively monitored cell membrane movements over 30 minutes to several hours with 500 nm lateral resolution using difference images.
    • Recorded faster cell movements within minutes using line scan mode.
    • Observed inhomogeneous volume changes across cell surfaces, suggesting the cytoskeleton's role in shape stabilization.

    Conclusions:

    • Pulse-mode SICM is a viable technique for quantitative analysis of cell volume and membrane dynamics during locomotion.
    • The method provides insights into the spatiotemporal regulation of cell shape and the influence of the cytoskeleton.
    • This imaging approach enables long-term, high-resolution studies of cellular mechanical processes.