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Quantifying cilia beat frequency using high-speed video microscopy: Assessing frame rate requirements when imaging

Luke Scopulovic1, Deanne Francis1, Elvis Pandzic2

  • 1Cilia Research Laboratory, College of Public Health Medical and Veterinary Sciences, James Cook University, Townsville, Queensland, Australia.

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|June 9, 2022
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Summary

Accurate cilia beat frequency (CBF) measurement requires video frame rates of at least 150 frames per second (fps). Higher rates offer minimal improvement, and the Nyquist theorem is unreliable for setting sampling rates for CBF assessment.

Keywords:
ciliacilia beat frequencyependymarespiratory epithelium

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Area of Science:

  • Biophysics
  • Cell Biology
  • Physiology

Background:

  • Motile cilia are crucial for physiological functions.
  • Cilia beat frequency (CBF) is a key metric for assessing cilia motility.
  • Inconsistent frame rates in video microscopy complicate CBF measurement.

Purpose of the Study:

  • Determine the optimal frame rate for accurate CBF assessment.
  • Evaluate the applicability of the Nyquist theorem for setting sampling rates.

Main Methods:

  • Collected high-speed videos (>600 fps) of mouse airway and ependymal cilia.
  • Down-sampled videos to various frame rates (30-300 fps).
  • Quantified CBF using manual counting and automated MATLAB scripts.

Main Results:

  • CBF measurements were significantly impaired at lower frame rates (30-60 fps).
  • Accurate CBF assessment required a minimum frame rate of 150 fps.
  • Automated CBF measurement accuracy improved at higher frame rates (>150 fps).

Conclusions:

  • The Nyquist theorem is not suitable for determining sampling rates for CBF measurement.
  • A sampling rate 3-4 times the CBF is recommended for accurate assessment, especially for automated methods.