We present a robot that enables high-content studies of alert adult Drosophila by combining operations including gentle picking; translations and rotations; characterizations of fly phenotypes and behaviors; microdissection; or release. To illustrate, we assessed fly morphology, tracked odor-evoked locomotion, sorted flies by sex, and dissected the cuticle to image neural activity. The robot's tireless capacity for precise manipulations enables a scalable platform for screening flies' complex attributes and behavioral patterns.
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We thank T.R. Clandinin, L. Liang, L. Luo, G. Dietzl, S. Sinha and T.M. Baer for advice; A. Janjua and J. Li for technical assistance; and D. Petrov and A. Bergland for providing inbred fly lines. We thank the W.M. Keck Foundation, the Stanford Bio-X program, a US National Institutes of Health Director's Pioneer Award for research funding (M.J.S.), and the Stanford-NIBIB Training program in Biomedical Imaging Instrumentation (J.R.M.).
Integrated supplementary information
The robot tracks and captures a fly under infrared illumination using real-time machine vision guidance.
Automated tracking and picking of an active fly from the picking platform, played back at real speed. Video 2 shows the same events in slow motion.
The same events as in Video 1, but played back at 20 × slower speed. To search for the ring reflection pattern on the fly thorax while tracking the fly, the robot turns on the ring of infrared LEDs before acquiring an image via the onboard camera. The robot picks up the fly by touching the picking effector to the reflected ring target on the thorax.
The picking platform is a key tool to enhance the throughput of automated handling. Alert flies that have never been anesthetized can rapidly populate the platform. These flies emerge through an opening in the center of the platform from a standard vial attached to the platform's underside.
After picking each fly, the robot carried it to an inspection camera and obtained the location of the neck apse. The robot used this information to align and tether the head of the fly to the holder. Since the head holders were based on suction, we released all three flies after the experiment.
The robot rotates a fly over 360° for high-magnification inspection at various yaw angles.
The robot rapidly transfers flies back and forth between the halves of a divided platform. This demonstration illustrates high-speed handling of flies.
The robot can pick and release individual flies multiple times without harming them.
To perform the discrimination, the robot picked individual flies and brought them to a high-magnification inspection camera. The system puffed air beneath the picked fly to induce flight, so that the wings did not occlude the abdomen. A robotic algorithm examined the fly as it was rotated, to find the best view of the abdomen, and then determined the fly's sex using an image of the abdomen (Supplementary Fig. 5 and Supplementary Table 2).
We recorded the forward, lateral, and rotational components of the locomotor patterns in response to odor stimuli, which we delivered through the pipette directed toward the fly's head.
Using a three-dimensional translation stage, we maneuvered the head-fixed fly to the end mill and made an initial cut to open the cuticle. Thereafter, we commanded the stage to continue cutting along a preprogrammed trajectory. Saline immersion kept the brain hydrated and prevented tissue debris from interfering with the surgery.
About this article
Nature Biotechnology (2017)