A MICROFLUIDIC CHIP FOR IMMOBILIZING AND IN VIVO IMAGING OF DROSOPHILA LARVAE

摘要

We present a 2-layer microfluidic PDMS chip for the long-term immobilization and in vivo imaging of Drosophila larvae. Using a combined CO_2/mechanical immobilization mechanism, we achieved high on-chip survival rates (up to 85% during a 10-hour time course) while we eliminated body movement which is required for high resolution imaging. We demonstrated the use of this technique for performing time-lapse imaging of axonal sprouting over a 12-hour period after laser axotomy. The noninvasive nature of the proposed method enabled for the first time to study in vivo axonal regeneration after laser injury. Our results suggest that the proximal stump of the injured axon remains dormant for 7 hours before regeneration is initiated.