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Multiphoton imaging of neural structure and activity in Drosophila through the intact cuticle

Summary

Scientists developed a new imaging technique that allows researchers to observe brain activity in fruit flies without surgically removing the protective head covering. This breakthrough lets researchers watch neural activity for much longer periods and during natural behaviors like walking and responding to odors. The technique uses special microscopes that shine infrared light through the fly’s intact head to image neurons expressing fluorescent proteins.

Background

Current fly neural imaging requires surgical removal of the head cuticle, which limits imaging duration due to brain tissue degeneration from damaged circulation. Long-term chronic recordings of neural activity are essential for understanding how neural circuits evaluate sensory information during different behavioral states.

Objective

To develop a multiphoton imaging method that enables neural structure and activity recording in behaving flies through the intact head cuticle, allowing extended imaging duration compared to current cuticle-removed preparations.

Results

The fly head cuticle showed high ballistic transmission (>90% at 1300 nm), with air sacs and underlying tissue being the primary impedance to imaging. Three-photon imaging outperformed two-photon imaging in deeper brain regions such as the central complex. Two-photon through-cuticle functional imaging successfully captured odor-evoked responses from mushroom body γ-lobes in behaving flies for up to 12 consecutive hours.

Conclusion

The cuticle-intact imaging method enables multiphoton imaging of fly neural structure and activity at long time scales without traumatic head surgery. This approach extends imaging duration capabilities and opens new possibilities for chronic neural recordings across the fly lifespan and multiple days of observation.
  • Published in:eLife,
  • Study Type:Technical Development Study,
  • Source: 10.7554/eLife.69094, 35073257
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