Medical & Biological Engineering & Computing

, Volume 53, Issue 10, pp 1025–1035

Robust estimation of the motile cilia beating frequency

Original Article

Abstract

The estimation of the cilia beating frequency (CBF) is of great interest in understanding how the CBF modulates liquid fluxes and how it is controlled by the ciliated cell intra- and/or extracellular medium composition in physiological processes. Motion artifacts and camera defaults may hinder the computation of the frequency variations during long-lasting experiments. We have developed a new analysis approach consisting of a preliminary corrective step (removal of a grid pattern on the image sequence and shift compensation), followed by a harmonic model of the observed cilia using a maximum likelihood estimator framework. It is shown that a more accurate estimation of the frequency can be obtained by averaging the squared Fourier transform of individual pixels followed by a particular summation over the different frequencies, namely the compressed spectrum. Robustness of the proposed method over traditional approaches is shown by several examples and simulations. The method is then applied to images of samples containing ciliated ependymal cells located in the third cerebral ventricle of mouse brains, showing that even small variations in CBF in response to changes in the amount of oxygenation, pH or glucose were clearly visible in the computed frequencies. As a conclusion, this method reveals a fine metabolic tuning of the cilia beating in ependimocytes lining the third cerebral ventricle. Such regulations are likely to participate in homeostatic mechanisms regulating CSF movements and brain energy supply.

Keywords

Signal processing Cilia Frequency estimation 

References

  1. 1.
    Chilvers MA, O’Callaghan C (2000) Analysis of ciliary beat pattern and beat frequency using digital high speed imaging: comparison with the photomultiplier and photodiode methods. Thorax 55:314–317PubMedCentralCrossRefPubMedGoogle Scholar
  2. 2.
    Conductier G, Brau F, Viola A, Langlet F, Ramkumar N, Dehouck B, Lemaire T, Chapot R, Lucas L, Rovère C, Maitre P, Hosseiny S, Petit-Paitel A, Adamantidis A, Lakaye B, Risold PY, Prévot V, Meste O, Nahon JL, Guyon A (2013) Melanin-concentrating hormone regulates beat frequency of ependymal cilia and ventricular volume. Nat Neurosci. doi:10.1038/NN.3401 PubMedGoogle Scholar
  3. 3.
    Del Bigio MR (2010) Ependymal cells: biology and pathology. Acta Neuropathol 119:55–73CrossRefPubMedGoogle Scholar
  4. 4.
    Dimova S, Maes F, Brewster ME, Jorissen M, Noppe M, Augustijns P (2005) High-speed digital imaging method for ciliary beat frequency measurement. J Pharm Pharmacol 57:521–526CrossRefPubMedGoogle Scholar
  5. 5.
    Garcia MA et al (2003) Hypothalamic ependymal–glial cells express the glucose transporter GLUT2, a protein involved in glucose sensing. J Neurochem 86:709–724CrossRefPubMedGoogle Scholar
  6. 6.
    Kay SM (1993) Fundamentals of statistical signal processing. Prentice Hall, Englewood CliffsGoogle Scholar
  7. 7.
    Kim W, Han TH, Kim HJ, Park MY, Kim KS, Park RW (2011) An automated measurement of ciliary beating frequency using a combined optical flow and peak detection. Health Inform Res 17(2):111–119CrossRefGoogle Scholar
  8. 8.
    Lechtrek KF, Delmotte P, Robinson ML, Sanderson MJ, Witman GB (2008) Mutations in Hydin impair ciliary motility in mice. J Cell Biol 180(3):633–643CrossRefGoogle Scholar
  9. 9.
    Lechtrek KF, Sanderson MJ, Witman GB (2009) High-speed digital imaging of ependymal cilia in the murine brain. Methods Cell Biol 91:255–264CrossRefGoogle Scholar
  10. 10.
    Liu L et al (2013) Method for quantitative study of airway functional microanatomy using micro-optical coherence tomography. PLoS One 8(1):e54473. doi:10.1371/journal.pone.0054473 PubMedCentralCrossRefPubMedGoogle Scholar
  11. 11.
    O’Callaghan C et al (2012) Analysis of ependymal ciliary beat pattern and beat frequency using high speed imaging: comparison with the photomultiplier and photodiode methods. Cilia 1(1):8PubMedCentralCrossRefPubMedGoogle Scholar
  12. 12.
    Oldenburg AL, Chhetri RK, Hill DB, Button B (2012) Monitoring airway mucus flow and ciliary activity with optical coherence tomography. Biomed Opt Express 3(9):1978–1992PubMedCentralCrossRefPubMedGoogle Scholar
  13. 13.
    Olm MA, Kogler JE, Macchione M, Shoemark A, Saldiva PH, Rodrigues JC (2011) Primary ciliary dyskinesia: evaluation using cilia beat frequency assessment via spectral analysis of digital microscopy images. J Appl Physiol 111:295–302PubMedCentralCrossRefPubMedGoogle Scholar
  14. 14.
    Psarra AM et al (1998) Immunocytochemical localization of glycogen phosphorylase kinase in rat brain sections and in glial and neuronal primary cultures. J Neurocytol 27:779–790CrossRefPubMedGoogle Scholar
  15. 15.
    Salathe M (2007) Regulation of mammalian ciliary beating. Annu Rev Physiol 69:401–422CrossRefPubMedGoogle Scholar
  16. 16.
    Schoberl M, Fossel S, Kaup A (2010) Fixed pattern noise column drift compensation (CDC) for digital moving picture camera. In: ICIP, pp 573–576Google Scholar
  17. 17.
    Smith CM, Djakow J, Free RC, Djakow P, Lonnen R, Williams G, Pohunek P, Hirst RA, Easton AJ, Andrew PW, O’Callaghan C (2012) CiliaFA: a research tool for automated, high-throughput measurement of ciliary beat frequency using freely available software. Cilia 1:14PubMedCentralCrossRefPubMedGoogle Scholar
  18. 18.
    Teff Z, Priel Z, Gheber LA (2008) The forces applied by cilia depend linearly on their frequency due to constant geometry of the effective stroke. Biophys J 94:298–305PubMedCentralCrossRefPubMedGoogle Scholar
  19. 19.
    Yi WJ, Park KS, Min YG, Sung MW (1997) Distribution mapping of ciliary beat frequencies of respiratory epithelium cells using image processing. Med Biol Eng Comput 35(6):595–599CrossRefPubMedGoogle Scholar
  20. 20.
    Yi WJ, Park KS, Lee CH, Rhee CS, Nam SW (2002) Directional disorder of ciliary metachronal waves using two-dimensional correlation map. IEEE Trans Biomed Eng 49(3):269–273CrossRefPubMedGoogle Scholar
  21. 21.
    Yi WJ, Park KS, Lee CH, Rhee CS (2003) Correlation between ciliary beat frequency and metachronal wave disorder using image analysis method. Med Biol Eng Comput 41(4):481–485CrossRefPubMedGoogle Scholar
  22. 22.
    Zitova B, Flusser J (2003) Image registration methods: a survey. Image Vis Comput 21(11):977–1000CrossRefGoogle Scholar

Copyright information

© International Federation for Medical and Biological Engineering 2015

Authors and Affiliations

  1. 1.Lab I3S, UMR 7271, CNRSUniversity of Nice-Sophia AntipolisSophia AntipolisFrance
  2. 2.IPMC, UMR 7275, CNRSUniversity of Nice-Sophia AntipolisValbonneFrance

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