A Nanosensor Toolbox for Rapid, Label-Free Measurement of Airway Surface Liquid and Epithelial Cell Function

Abstract

Ciliated lung epithelial cells and the airway surface liquid (ASL) comprise one of the body's most important protective systems. This system is finely tuned, and perturbations to ASL rheology, ASL depth, ASL pH, the transepithelial potential, and the cilia beat frequency are all associated with disease pathology. Further, these apparently distinct properties interact with each other in a complex manner. For example, changes in ASL rheology can result from altered mucin secretion, changes in ASL pH, or changes in ASL depth. Thus, one of the great challenges in trying to understand airway pathology is that the properties of the ASL/epithelial cell system need to be assessed nearsimultaneously and without perturbing the sample. Here, we show that nanosensor probes mounted on a scanning ion conductance microscope make this possible for the first time, without any need for labeling. We also demonstrate that ASL from senescenceretarded human bronchial epithelial cells retains its native properties. Our results demonstrate that by using a nanosensor approach, it is possible to pursue faster, more accurate, more coherent, and more informative studies of ASL and airway epithelia in health and disease.

Applications of scanning ion conductance microscope in biomedical fields

“…Ivanova et al realized the evaluation of the interference‐free characteristics of the ciliated lung epithelial cells and airway surface fluid (ASL)/epithelial cell system for the first time through the nanosensor probes mounted on a scanning ion conductance microscope. It is of great importance to conduct faster, more accurate, more consistent and more informative research on ASL and airway epithelium in health or disease 88 . Therefore, the use of SICM observation of organizational structure is helpful to reveal the mechanism of various diseases and to identify potential treatment strategies.…”

“…It is of great importance to conduct faster, more accurate, more consistent and more informative research on ASL and airway epithelium in health or disease. 88 Therefore, the use of SICM observation of organizational structure is helpful to reveal the mechanism of various diseases and to identify potential treatment strategies.…”

Application of scanning ion conductance microscope in cell characterizations

“…[57] ; (b) 活的海马神经元高分辨率SICM 成像 [19] ; (c) 单个纳米粒子与细胞膜相互作用动态过程观察的荧光和SICM形貌图 [60] ; (d) 玉米根毛细胞对[Ru(NH 3 ) 6 ] 3+ 的摄入 过程的SICM和SECM图像 [63] (网络版彩图) 呈现细胞表面形态及纳米颗粒内吞早期阶段的动态交 替过程. Su等 [62] [38] 、 心肌细胞 [39] 、上皮细胞 [30,69,70] 、成纤维细胞 [37,45] 、神 经元细胞 [32,71~74] 和胶质细胞 [75] 等. 例如, Happel等 [38] 使 用SICM观察到了在高渗和低渗溶液中肝细胞体积的 调节过程.…”

Research on the Adaptive Sensitivity Scanning Method for Ion Conductance Microscopy with High Efficiency and Reliability

“…Scanning ion conductance microscopy (SICM) is an emerging scanning probe technique that features an in situ noncontact measurement capability with nanoscale resolution and is used increasingly for sample observation in fields including cytobiology, , pathology, , electrochemistry, , and material science. , The principle of SICM is that the ion current flow through the probe tip, a pipet with a micro- or nanoscale tip opening, is monitored to act as a feedback signal to maintain noncontact sample scanning . This feedback mechanism allows SICM to have strong adaptability to samples with different conductivities and stiffnesses. , During SICM scanning, the detectable pipet–sample distance is called the sensitivity and is a key parameter in determining the scanning speed and accuracy of the pipet .…”