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Related Experiment Video

Updated: May 21, 2026

Longitudinal Measurement of Extracellular Matrix Rigidity in 3D Tumor Models Using Particle-tracking Microrheology
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Published on: June 10, 2014

Depth-resolved cellular microrheology using HiLo microscopy.

Jarett Michaelson, Heejin Choi, Peter So

    Biomedical Optics Express
    |June 29, 2012
    PubMed
    Summary
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    Accurate cell mechanical property measurements require removing out-of-plane data. New HiLo microscopy techniques significantly improve viscoelastic property analysis in 3D cell environments.

    Area of Science:

    • Biophysics
    • Cell Biology
    • Microscopy

    Background:

    • Measuring cell mechanical properties in 3D is crucial.
    • Conventional particle tracking microrheology (PTM) can be affected by out-of-plane artifacts.

    Purpose of the Study:

    • To develop and validate a microscopy technique to reduce out-of-plane contributions in PTM.
    • To accurately assess cell viscoelastic properties in 3D environments.

    Main Methods:

    • Utilized HiLo microscopy to selectively eliminate out-of-plane probe signals.
    • Developed a method to remove signals from probes 0.5 μm or further from the focal plane.
    • Applied the technique to characterize live-cell bilayers.

    Main Results:

    Keywords:
    (170.1530) Cell analysis(180.0180) Microscopy

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  • The HiLo microscopy technique effectively removed 90% of out-of-plane signals while retaining in-plane probes.
  • Significant, frequency-dependent differences in extracted cell moduli were observed compared to conventional analysis.
  • Demonstrated the impact of out-of-plane data removal on mechanical property assessment.

    • Conclusions:

    • Removal of out-of-plane information is vital for accurate cell mechanical property measurements.
    • HiLo microscopy offers an improved approach for PTM in 3D cellular environments.
    • This technique enhances the reliability of viscoelastic property analysis in live cells.