Real-Time 3-D Microscopy of Freely Moving <i>Drosophila melanogaster</i> Larvae (VIDEO)
Caption
This movie shows different visualizations of high-speed volumetric microscopy data acquired on transgenic Drosophila larvae. The movie showcases the ability of SCAPE to acquire structural and functional 3-D information in freely moving whole organisms in real-time.
The first SCAPE sequence shows 4 (out of 80) depths within a freely moving mhc-GFP 1st instar larva whose body muscles, heart and gut are labeled with GFP. The larva is freely crawling on a glass slide and is being imaged using a single, stationary objective lens at 20 volumes per second. The beating heart can be observed.
The second SCAPE sequence shows a similar 3rd instar larva co-expressing GCaMP and mCherry in muscle. The dynamics of muscle depolarization can be visualized from the calcium-sensitive GCaMP fluorescence, while the mCherry captures the 3D dynamics of motion. Data was acquired at 10 volumes per second.
The third SCAPE sequence shows whole body imaging of a 1stinstar larva of the same mCherry-GCaMP mhc strain. Different components of the data are rendered and visualized separately and together. The larva was attached to a glass slide on its ventral slide to prevent motion out of the field of view, but waves of peristaltic muscle contraction can be observed.
4th SCAPE sequence shows an adult Drosophila larva acquired with much lower magnification (after optical clearing). Additional published results show neuronal labeling and 4D tracking.
Credit
Elizabeth Hillman, Columbia Engineering
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