Ex vivo Method for High Resolution Imaging of Cilia Motility in Rodent Airway Epithelia

被引:38
作者
Francis, Richard [1 ]
Lo, Cecilia [1 ]
机构
[1] Univ Pittsburgh, Dept Dev Biol, Pittsburgh, PA 15260 USA
来源
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2013年 / 78期
关键词
Biomedical Engineering; Issue; 78; Developmental Biology; Cellular Biology; Molecular Biology; Anatomy; Physiology; Respiratory Mucosa; Trachea; Ciliary Motility Disorders; Animal Experimentation; Microscopy; Fluorescence; Interference; Polarization; Video; Airway; mucociliary clearance; microscopy; animal model;
D O I
10.3791/50343
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
An ex vivo technique for imaging mouse airway epithelia for quantitative analysis of motile cilia function important for insight into mucociliary clearance function has been established. Freshly harvested mouse trachea is cut longitudinally through the trachealis muscle and mounted in a shallow walled chamber on a glass-bottomed dish. The trachea sample is positioned along its long axis to take advantage of the trachealis muscle to curl longitudinally. This allows imaging of ciliary motion in the profile view along the entire tracheal length. Videos at 200 frames/sec are obtained using differential interference contrast microscopy and a high speed digital camera to allow quantitative analysis of cilia beat frequency and ciliary waveform. With the addition of fluorescent beads during imaging, cilia generated fluid flow also can be determined. The protocol time spans approximately 30 min, with 5 min for chamber preparation, 5-10 min for sample mounting, and 10-15 min for videomicroscopy.
引用
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页数:6
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