Figure 2.

Whole animal Ca²⁺-imaging of Caenorhabditis elegans using light-fieldmicroscopy. (a) Wide-field image of the worm inside a microfluidicpoly(dimethylsiloxane) (PDMS) device used to immobilize the worm. Head is atbottom right. (b) Maximum intensity projection (MIP) of light-field deconvolvedimage (15 iterations) containing 14 distinct z-planes. The majority of the wormand its nervous system are clearly visible, including the head ganglia (at bottomright) as well as the ventral cord. Scale bar 50 μm. (c) Ca²⁺-intensity traces(ΔF/F₀) of NLS-GCaMP5K fluorescence of selected neurons in the ventral cordand head region, as indicated by colored arrows and numbers in (b), and imagedvolumetrically at 5 Hz for 200 seconds. Also see Supplementary Video 1. (d)Zoom of the brain region, with MIP of xy plane as well as xz and yz cross-sections indicated by the dashed lines. Single neurons are well resolved (Supplementary Video 2). Scale bar 10 μm. (e) Individual z planes of typical recording of theworm's brain, reconstructed from a single camera exposure (see also Supplementary Fig. 2 for neuron IDs). In this recording, the worm's center wasplaced at the focal plane of the objective. Scale bar 50 μm. (f) Activity of all 74neurons identified in (e), also see Supplementary Video 4. Each row shows atime-series heat map. Color indicates percent fluorescence changes (ΔF/F₀);scaling is indicated by the color bar on the right. The x-axis in (f) representselapsed recording time while the y axis shows individual neurons.