Home › Cell Biology › Using Microfluidics Chips for Live Imaging and Study of Injury Responses in Drosophila Larvae
Cell Biology JoVE (Open Access) Citable · DOI

Using Microfluidics Chips for Live Imaging and Study of Injury Responses in Drosophila Larvae

DOI: 10.3791/50998-v
What you'll learn
  • ✓ Fabricate and prepare single-layer PDMS microfluidic devices for larval imaging
  • ✓ Perform live-cell imaging of neuronal processes in intact Drosophila larvae
  • ✓ Induce and visualize nerve crush injuries in larval segmental nerves
  • ✓ Apply immobilization techniques to enhance image quality during dynamic imaging
Protocol

Drosophila larvae are an attractive model system for live imaging due to their translucent cuticle and powerful genetics. This protocol describes how to utilize a single-layer PDMS device, called the 'larva chip' for live imaging of cellular processes within neurons of 3rd instar Drosophila larvae.

Difficulty
intermediate
Total time
~3-4 hours per experiment (chip prep ~1-2 hrs, imaging/injury induction ~2 hrs)
Model organism
Drosophila melanogaster (3rd instar larvae)

Steps

1
Prepare single-layer PDMS microfluidic chip

Fabricate and cure a PDMS device (larva chip) from mold using standard soft lithography. Prepare the chip surface for larval mounting and imaging.

▶ 01:47
2
Mount larva and perform live imaging

Position 3rd instar larva within the chip device and acquire time-lapse images of neuronal cellular processes using fluorescence microscopy.

▶ 03:47
3
Induce nerve crush injury in segmental nerves

Apply mechanical crush injury to larval segmental nerves using micromanipulation while larva remains in the imaging chamber, enabling real-time visualization of injury response.

▶ 07:23
4
Enhance image quality with immobilization

Apply immobilization techniques to reduce larval movement artifacts and improve signal clarity during extended live-imaging sessions of injury responses.

▶ 08:42
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