Schipor Ioana, Palmer James N, Cohen Akiva S, Cohen Noam A
Department of Otorhinolaryngology-Head and Neck Surgery, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
Am J Rhinol. 2006 Jan-Feb;20(1):124-7.
Mucociliary clearance is a critical upper airway host defense mechanism. Ciliated epithelium in the mammalian airway continually beat at a baseline frequency. Importantly, during times of stress such as exercise or infection, the cilia beat faster to increase clearance. Nasal epithelial ciliary beat frequency (CBF) has been analyzed previously in ex vivo specimens using a variety of methods including photodiode detectors and conventional video recording. Recent studies performed using lower airway ciliated mucosa have shown poor correlation between CBF quantified by photodiode/conventional video and those using high-speed digital video capture at temperatures close to physiological temperatures. Thus, to more rigorously interrogate sinonasal CBF at physiological conditions, we have incorporated a high-speed digital video camera to our CBF analysis system. This is the first report of sinonasal epithelial CBF analysis performed using high-speed video digital analysis.
Ex vivo samples of sinonasal epithelium were placed in lactated Ringer's in a temperature-controlled microscope stage chamber. An edge of tissue containing beating cilia was observed at a magnification of 630x using differential interference contrast microscopy. The images were captured using a high-speed digital camera with a sampling rate of 250 frames per second. CBF was determined using computerized data analysis.
The mean nasal CBF was calculated from a minimum of five regions for each sample. Temperature curves were generated from tissue obtained from chronically infected subjects.
Analysis of high-speed digital video capture of sinonasal CBF observed under differential interference contrast microscopy is a powerful method to investigate environmental as well as host influences on mucociliary clearance within the upper airways.
黏液纤毛清除是上呼吸道重要的宿主防御机制。哺乳动物气道中的纤毛上皮细胞持续以基线频率摆动。重要的是，在运动或感染等应激状态下，纤毛摆动加快以增加清除率。此前已使用包括光电二极管探测器和传统视频记录在内的多种方法，对离体标本中的鼻上皮纤毛摆动频率（CBF）进行了分析。最近使用下呼吸道纤毛黏膜进行的研究表明，在接近生理温度的条件下，通过光电二极管/传统视频量化的CBF与使用高速数字视频捕获量化的CBF之间相关性较差。因此，为了在生理条件下更严格地研究鼻窦CBF，我们在CBF分析系统中加入了高速数字摄像机。这是首次使用高速视频数字分析进行鼻窦上皮CBF分析的报告。
将鼻窦上皮的离体样本置于温度可控的显微镜载物台腔室中的乳酸林格液中。使用微分干涉相差显微镜以630倍放大率观察含有摆动纤毛的组织边缘。使用采样率为每秒250帧的高速数字相机捕获图像。使用计算机数据分析确定CBF。
计算每个样本至少五个区域的平均鼻CBF。从慢性感染受试者获得的组织生成温度曲线。
在微分干涉相差显微镜下观察到的鼻窦CBF的高速数字视频捕获分析，是一种研究环境以及宿主对上呼吸道黏液纤毛清除影响的有力方法。
