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David Deleted File
about 8 years ago
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Here we describe an affordable and scalable method for automatically assaying pharyngeal pumping. Our method combines *our* previously described microfluidic device \cite{KopitoLabChip}, low cost educational microscopes, and an image analysis pipeline implemented using top tier open source tools and libraries, among the most widely used by researchers. The advantages of our approach include precise control of conditions such as the quality, uniformity, and concentration of available food, the possibility of prolonged measurement durations (hours, if required), unbiased automatic detection of pumping events, and the possibility to assay feeding conditions that change dynamically in a controlled manner.
Manual scoring of pumping is arduous and limited to brief measurement periods. Automatic scoring of pumping on a high quality microscope is throughput-limited due to the cost of the imaging setup. We found that lower quality imaging can be compensated for by rapid sampling and improved analysis without compromising the quality of the data. Using three microscopes we were able to assay up to 50 animals a day, i.e., 6 animals per objective, each for a full hour, at the approximate cost of $2,000 (not including optional syringe pumps). The setup can be further duplicated and the rate limiting step is number of microfluidic devices that the researcher can load. The flexibility dynamically controlling the feeding conditions is dictated by the sophistication of the pressure source that is driving the fluid flow.
