Ciliary Beat of Cultured Human Respiratory Cells Studied with Differential Interference Microscope and High Speed Video System

Rautiainen, Markus; Matsune, Shouji; Shima, Sota; Sakamoto, Kunihiko; Hanamure, Yutaka; Ohyama, Masaru

doi:10.3109/00016489209137483

Cited by 23 publications

(15 citation statements)

References 10 publications

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“…Normal values of the different parameters according to the literature data for B-SIT score,18 TEM analysis of cilia,19 amount of ciliated cells and ciliated edges with beating cilia,20 cilia length,21 ciliary beat frequency22 and beating angle 23. The value of the expected running distance per second was calculated using the following formula: length×angle×frequency×1 s.…”

Section: Methodsmentioning

confidence: 99%

Abnormal respiratory cilia in non-syndromic Leber congenital amaurosis with CEP290 mutations

Papon

Perrault

Coste

et al. 2010

Journal of Medical Genetics

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These data provide the first clear demonstration of respiratory cilia ultrastructural defects in LCA patients with CEP290 mutations. The frequency of these findings in LCA patients along with the high expression of CEP290 in nasal epithelium suggest that CEP290 has an important role in the proper development of both the respiratory ciliary structures and the connecting cilia of photoreceptors. The presence of respiratory symptoms in patients could represent additional clinical criteria to direct CEP290 genotyping of patients affected with the genetically heterogeneous cone-rod dystrophy subtype of LCA.

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Section: Methodsmentioning

confidence: 99%

Abnormal respiratory cilia in non-syndromic Leber congenital amaurosis with CEP290 mutations

Papon

Perrault

Coste

et al. 2010

Journal of Medical Genetics

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“…CBF data can also be obtained by monitoring ciliary beating using a video camera or a digital high-speed video system, followed by off-line analysis either using manual counting of the ciliary beats on replaying the pictures over defined time intervals (Rautiainen et al 1992;Chilvers & O'Callaghan 2000;Doran et al 2004) or by analysing changes in image intensity at defined locations (Nasr et al 1995;Sanderson 2000). The use of standard video camera equipment induces a clinically relevant upper limit of measurable frequencies, while the use of a highspeed camera is time-consuming and reduces the number of cilia measured in one sample.…”

Section: Introductionmentioning

confidence: 99%

High-speed digital imaging method for ciliary beat frequency measurement

Dimova

Maes

Brewster

et al. 2005

Journal of Pharmacy and Pharmacology

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The aim of this study was to develop a high-speed digital imaging system and related software for ciliary beat frequency (CFB) analysis in order to establish an automated and reliable method that is observer independent and faster compared to the conventional computerized microscope photometry method. Using primary human nasal epithelial cell cultures, the CBF was recorded with a computerized microscope photometry system and a high-speed digital imaging system. To obtain a wide range of frequencies, glycocholate (0.5%) and chlorocresol (0.005%) were used as ciliostimulatory and cilio-inhibitory reference compounds, respectively. The mean values in hertz (+/- s.d.) obtained with the photometry and high-speed digital imaging systems were: controls 8.2 +/- 0.9 and 7.9 +/- 1.1; chlorocresol 5.0 +/- 0.9 and 5.1 +/- 1.1; glycocholate 9.8 +/- 1.0 and 9.7 +/- 0.8. A similar increase (by 20 and 24%) and decrease (by 38 and 35%) in CBF was determined by the two methods after glycocholate and chlorocresol treatment, respectively. The mean difference between the photometry and high-speed digital imaging methods was 0.2 +/- 0.6 Hz, and the Bland-Altman limits of agreement were from -1.0 to +1.4 Hz, suggesting that the results obtained by these two methods could be used interchangeably. These results show the reliability of the high-speed digital imaging system and the software developed for in-vitro CBF measurements. The advantages of the system include: (i) fast data acquisition and calculation, (ii) whole field automated CBF analysis and (iii) reduction in selection bias.

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“…Over recent years, high speed digital video cameras have been developed, allowing recordings to be made at frame rates up to 45,000 per second. High speed digital video cameras provide a simple and accurate method for determining ciliary beat frequency [25] and beat pattern of individual cilia may be seen and analyzed in slow motion following recording [19,20,26]. There have been no direct comparisons of the use of the photodiode, photomultiplier and high speed video techniques to measure the beat frequency of ependymal cilia.…”

Section: Introductionmentioning

confidence: 99%

Analysis of ependymal ciliary beat pattern and beat frequency using high speed imaging: comparison with the photomultiplier and photodiode methods

O’Callaghan

Sikand

Chilvers

2012

Cilia

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BackgroundThe aim of this study was to compare beat frequency measurements of ependymal cilia made by digital high speed imaging to those obtained using the photomultiplier and modified photodiode techniques. Using high speed video analysis the relationship of the power and recover strokes was also determined.MethodsCiliated strips of ependyma attached to slices from the brain of Wistar rats were incubated at 30°C and observed using a ×50 water immersion lens. Ciliary beat frequency was measured using each of the three techniques: the high speed video, photodiode and photomultiplier. Readings were repeated after 30 minutes incubation at 37°C. Ependymal cilia were observed in slow motion and the precise movement of cilia during the recovery stroke relative to the path travelled during the power stroke was measured.ResultsThe mean (95% confidence intervals) beat frequencies determined by the high speed video, photomultiplier and photodiode at 30°C were 27.7 (26.6 to 28.8), 25.5 (24.4 to 26.6) and 20.8 (20.4 to 21.3) Hz, respectively. The mean (95% confidence intervals) beat frequencies determined by the high speed video, photomultiplier and photodiode at 37°C were 36.4 (34 to 39.5), 38.4 (36.8 to 39.9) and 18.8 (16.9 to 20.5) Hz. The inter and intra observer reliability for measurement of ciliary beat frequency was 3.8% and 1%, respectively. Ependymal cilia were observed to move in a planar fashion during the power and recovery strokes with a maximum deviation to the right of the midline of 12.1(11.8 to 13.0)° during the power stroke and 12.6(11.6 to 13.6)° to the left of the midline during the recovery stroke.ConclusionThe photodiode technique greatly underestimates ciliary beat frequency and should not be used to measure ependymal ciliary beat frequency at the temperatures studied. Ciliary beat frequency from the high speed video and photomultiplier techniques cannot be used interchangeably. Ependymal cilia had minimal deviation to the right side during their power stroke and to the left during the recovery stroke.

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Ciliary Beat of Cultured Human Respiratory Cells Studied with Differential Interference Microscope and High Speed Video System

Cited by 23 publications

References 10 publications

Abnormal respiratory cilia in non-syndromic Leber congenital amaurosis with CEP290 mutations

Abnormal respiratory cilia in non-syndromic Leber congenital amaurosis with CEP290 mutations

High-speed digital imaging method for ciliary beat frequency measurement

Analysis of ependymal ciliary beat pattern and beat frequency using high speed imaging: comparison with the photomultiplier and photodiode methods

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