Fig 1.
Dynamic photopatterning method and implementation.
(a) Sequence, including real-time image capture and decision-based projection of the next fabrication step. (b) Schematic of the dynamic photopatterning system and configuration for patterning agar plates during C. elegans culture. Photo of the system is shown in S1 Fig. (c) An example of a framed micropillar array formed around a C. elegans worm in situ.
Fig 2.
Resolution and exposure data for PEG-DA on agar.
(a) Optical images of fabricated test pattern (USAF 1951 pattern) used to determine line pair resolution, with examples (from left to right) of under-exposed, properly exposed, and over-exposed results. (b) Relationship between line pair resolution and feature height for different objective magnifications indicated in the legend. (c) Relationship between optimal exposure time and feature height, determined using smallest line pair fabricated in each case. (d) SEM images of T-mazes as used in Fig 6, including (from left to right) the full maze in the upper half and the projected mask overlaid in the lower half, the sub-image shows one leg of the maze, and the right sub-image shows the corner of a wall feature.
Fig 3.
In situ photopolymerization of microstructures resulting in physical confinement of a C. elegans.
(a) Three exemplary assays built sequentially (from left to right): an open frame, an array of micropillars (100 μm diameter), and a rippled microchannel (approx. 200 μm wide). (b) Tracking of the worm motion over a time period of 200s, within the pillar array. (c) Box-whisker plots of velocity in each configuration, showing that sequential confinement increases the maximum velocity at which the worm pushes against the surface of features. S1 Video shows the experiment.
Fig 4.
Interaction between worms and a simple hinge-pin mechanism fabricated within a millimeter-scale area.
(a) Sequential frames show the lower worm contacting the hinge and extending its body, causing the hinge to rotate around the encapsulated pin. (b) Angle of the hinge with respect to vertical position plotted against time. Over six seconds the hinge rotates from 6° to 27°. (c) Schematics of worm motion as observed in the experiment, which is also shown in S2 Video. When the worm contacts two points spanning from the frame to the hinge, it extends its body, exerting force on the hinge and causing it to rotate. The motion cycle is repeated, rotating the hinge clockwise a small amount with each cycle.
Fig 5.
Tablet-based method for real-time assay modification by hand drawing.
(a) Schematic of sequence, resulting in projection of manual tablet input to create PEG-DA features on the substrate. In this case, the scale of the hand drawing is reduced 50X. (b) Video frames of the photopatterning of a series of dots drawn by the researcher to confine a worm inside a spiral frame (S3 Video). The system projects each feature drawn by the researcher after a delay of 0.25 seconds.
Fig 6.
Fabrication of maze assays as a test case of C. elegans decision-making behavior.
(a) Array of T-shaped mazes. The observed ripples are part of the agar surface. (b) T-shaped PEG-DA microchannel, with dot sequence indicating the centroid position of the worm during a 30 second period. (c) Percentage of worms that ended at each leg of the maze after being inserted into the entrance of the maze. See S4 Video for video of a nematode solving a maze.