This page lists techniques and measurements employed by the physioLens system
The physioLens is equipped with one brightfield LED and has the option of having three fluorescence LEDs. The LED used to excite the channel can be selected instantaneously.
Repeated imaging of multiple airways is accomplished by automated XY stage with micron accuracy thanks to feedback-controlled positioning. A low magnification objective (4x) allows for scanning six entire slices in under 10 minutes. The compiled images allow scientists to find airways in an intuitive manner.
The physioLens currently provides the lumen area of tissue slices, which in turn is used to calculate the bronchoconstriction of the airways of sizes ranging from 50 to over 500 microns. Automated lumen area measurements are accomplished by edge detection algorithms.
The physioLens accurately measures cilia beating frequency within the physioWare software. It provides results as frequency maps and averages, aligning well with known values from published literature. The system is optimized to measure frequencies up to 25 beats per second, with the potential for even higher measurements at faster recording speeds.
Cilia beating within area of interest
A calcium oscillation analyzer converts fluorescence brightness fluctuations into oscillation frequencies thanks to the use of fluorescent calcium indicators.
Agonists are provided to the slice without user intervention by a dosing nozzle, that can provide a 95% media exchange in less than 20 seconds. In addition, eight dosing bottles are provided to complete a detailed dose response automatically. The dosing module may be used to wash slices or add different chemical agents.
