Compared with conventional confocal microscopy and other high-resolution imaging techniques, this approach is distinguished by its simplicity, cost-effectiveness, and ease of standardization. It eliminates the need for high-powered optics and complex sample preparation, enabling rapid, reproducible measurements across multiple wells using common laboratory equipment. Validated against confocal measurements, it shows strong correlation and reliability. Its compatibility with high-throughput screening makes it ideally suited for studying airway hydration dynamics, evaluating potential cystic fibrosis therapies, and assessing physiological responses of human bronchial epithelial cultures under various conditions.
Description
This method uses light refraction imaging to noninvasively measure airway surface liquid volume in epithelial cell cultures grown at an air–liquid interface. A light source illuminates each well to form a fluid meniscus around the perimeter, and a scanner or camera captures the refracted light as a grayscale intensity profile. Analysis software processes the image by comparing it to calibrated standards or fitting a mathematical model (for example, a four-parameter logistic function) to extract parameters like half-maximal effective concentration or area under the curve. An optically active composition such as a fluorescent dye or light-scattering particles can be added to enhance contrast. The complete setup consists of a well-plate assembly, an imaging device, and a computer running specialized software.
Applications
High-throughput drug screening assays
Cystic fibrosis therapeutic screening
Respiratory research assay platform
Airway hydration monitoring kits
Laboratory imaging instrumentation sales
Advantages
Simple, standardized measurement of ASL volume without high-powered microscopy
Non-invasive light-refraction imaging using common scanners or cameras
Enhanced contrast with fluorescent dyes or scattering particles for greater accuracy
Quantitative outputs (e.g., EC₅₀, AUC) for reproducible analysis
High-throughput compatibility for screening airway-targeted therapeutics
Cost-effective and widely accessible compared to confocal microscopy
Validated correlation with traditional confocal measurements
Direct applicability to cystic fibrosis and airway disease research
IP Status
https://patents.google.com/patent/US9784725B2
