呼吸道纤毛运动状况的高速数字化显微成像  

Measurement of respiratory ciliary beat frequency quantified with high-speed digital microscopy

作　　者：张罗[1] 韩德民[1] 王鸿[1] 周兵[1] Michael J.Sanderson 

机构地区：[1]首都医科大学附属北京同仁医院耳鼻咽喉-头颈外科 北京市耳鼻咽喉科研究所,北京100730 [2]Division of Biology University of Massachusetts Medical School

出　　处：《中华耳鼻咽喉科杂志》2004年第7期433-437,共5页Chinese Journal of Otorhinolaryngology

基　　金：北京市科技新星计划资助项目(2003-B-15)

摘　　要：目的 通过对呼吸道纤毛摆动的频率和形式进行精确测量,以建立客观评价呼吸道上皮纤毛传输功能的指标。方法 采用高速数字化显微成像技术,对体外培养的兔和鼠气管黏膜以及人鼻腔黏膜的纤毛摆动状况进行观察。结果 ①数字化图像中测量点灰度的变化反映纤毛运动的状况,通过分析图像灰度的变化过程,可评价纤毛运动周期中各个时段的特征变化;②图像采集系统的采样速率为240帧/s时,可清晰显示纤毛运动状况,而当采样速率下降为30帧/s时,则无法获得正确的纤毛摆动频率(ciliary beat frequency,CBF)数据;③30℃时,鼠呼吸道CBF基值(x±sx)为(15.6±0.7)Hz(n=5);兔呼吸道CBF基值为(13.2±0.9)Hz(n=7);人鼻腔CBF基值为(13.4±1.1)Hz(n=5),3者间差异无显著性(P>0.05)。当向培养液中加入ATP后,CBF将在数秒内快速升高。结论 高速数字化显微成像和精确的数据处理技术,能够准确反映呼吸道纤毛运动状况,为客观评价呼吸道黏液纤毛传输功能提供临床实用指标。Objective To accurately quantify the respiratory cilia activity, a high-speed phase-contrast imaging and ciliary beat frequency (CBF) analysis method were introduced. Methods Airway ciliated cells, from rabbit trachea, mouse trachea and human nose, were prepared by primary culture of airway epithelial cells. Phase-contrast images were detected with a digital high-speed camera, a progressive scan charge-coupled device(CCD) that provided images (648 pixels x 200 lines) at 240 frames per second (fps). A data analysis approach, which can measure the period [frequency (Hz) = 1/period] of each ciliary beat cycle to match the high temporal resolution of the image acquisition rate, was also introduced. Results The high signal-to-noise ratio of gray waveform was obtained by interactively selecting the optimal region of interest. The frequency of each ciliary beat cycle was determined from the period of each cycle of the gray -intensity waveform. At 30℃, the basal CBF (x ?sx) , from mouse, rabbit and human, were (15. 6 ?0. 7 ) Hz ( n = 5 ) , (13. 2 ?0. 9 ) Hz (n = 7 ) , and (13.4 ?1. 1) Hz ( re = 5) , respectively. No statistical difference was found among groups (P >0. 05 ). In response to extracellular ATP, CBF had a rapid increment that could occur in seconds, a behavior suggesting the necessity of the high temporary resolution data acquisition system. Conclusions This ease of recording and replaying high-speed images may be useful in analyzing different phases and forms of ciliary beat patterns and perhaps in the diagnosis of cilia dysfunction-related diseases.

关 键 词：呼吸道 纤毛运动 高速数字化显微成像 纤毛上皮细胞 诊断 

分 类 号：R310[医药卫生—基础医学]