Dual color acquisition

The contraction of the animal's skeletal and smooth muscles generates tissue motion, which can interfere with the analysis of calcium signals; as a result, temporal volumetric registration is necessary prior to the extraction of cellular activity traces. To aid alignment, we performed dual-color imaging of the calcium imaging channel (pancellular GCaMP7f) concurrently with an anatomical reference channel. For this reference channel, we generated a new panmembrane transgenic line, Tg(β-actin:mCherry-CAAX), which labels all cell membranes to provide high-spatial-frequency textures throughout the organism.

Example plane of anatomical reference channel (left) and physiological channel (right). The anatomical channel is based on a transgenic line what expresses a membrane-targeted fluorescent in all cells providing high-frequency texture across the body.