Paper BM+MN+BI+BO-TuM6
Automated on-Chip Rapid Microscopy, Phenotyping, and Screening of C. elegans

Tuesday, October 21, 2008, 9:40 am, Room 309

Microscopy, phenotyping, and visual screens are critical methods frequently applied to model organisms in combination with genetics. Although widely used, these techniques for multicellular organisms have mostly remained manual and low-throughput. We report the complete automation of sample handling, high-resolution microscopy, phenotyping, and screening of C. elegans using a custom-designed microfluidic system. The engineered system, coupled with customized software, enables high-throughput diffraction-limited imaging and sorting of samples with no human intervention with any microscopy setup. The robustness and automation of our system relies greatly on integrated closed-loop control software as well as engineered hardware design of the microchip. The chip has six salient features that ensure a consistent and reliable operation for an extended period of time. First, it automatically self-regulates the loading of nematodes by a simple passive loading-regulator design. Constant pressure drives the flow, so that no feedback or intervention is necessary for the microchip to allow one and only one animal to occupy the imaging area at a time. Second, the setup automatically positions the samples in an identical position in the chip, so as to minimize the travel of the motorized stage and thereby reduce the processing time and increase the throughput. Third, the device has an integrated local temperature control system whereby animals are cooled to ~4 °C and completely immobilized briefly (~ a few seconds) for imaging and manipulation without the use of anesthetic drugs. Cooling provides an alternative to anesthetics, potentially minimizing the adverse developmental effects. Fourth, the microchip and the setup are compatible with any standard microscopy setup with no modification necessary, including simple compound epifluorescence microscopy, as well as more expensive multiphoton or confocal microscopy. Fifth, the microchip has no permanent small features (<20 μm), and therefore is easy to fabricate, less likely to be clogged by debris, and can operate very robustly. Lastly, losses through our system are minimal (~3%,) and the device design is gentle on the animals as the viability of all the sorted animals is ~100%. We show that compared to standard manual operation, time for phenotyping and visual screens can be reduced by ~2 orders of magnitude in our system with no human intervention, which has not been demonstrated before. Moreover, we show the ability to perform multiple sensitive and quantitative genetic screens with real biological samples based on cellular and subcellular features with over 95% accuracy per round.