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Microscopy & Imaging JoVE (Open Access) Citable · DOI

Agarose Microchambers for Long-term Calcium Imaging of Caenorhabditis elegans

DOI: 10.3791/52742-v
What you'll learn
  • ✓ Construct agarose microfluidic chambers for C. elegans imaging
  • ✓ Perform long-term calcium imaging without animal immobilization
  • ✓ Acquire neural activity data across all C. elegans life stages
Protocol

Imaging behavior and neural activity over long time scales without immobilization of the animal is a prerequisite to understand behavior. Agarose microfluidic chambers imaging (AMI) can be used to image neural activity and behavior for all life stages of Caenorhabditis elegans.

Difficulty
intermediate
Total time
~2-3 hours per imaging session (chamber preparation ~1 hour, imaging acquisition variable)
Model organism
Caenorhabditis elegans
Biosafety
BSL-1

Steps

1
Assemble imaging instruments and culture media

Gather required instruments, culture media, and petri dishes for agarose chamber preparation and C. elegans maintenance. Verify all equipment is functional before proceeding.

▶ 01:00
2
Prepare agarose microchamber device

Fabricate agarose microfluidic chambers following established protocols. This involves casting agarose into molds to create discrete chambers for individual C. elegans.

▶ 01:58
3
Load worms into microfluidic imaging chambers

Transfer C. elegans specimens into prepared agarose microchambers and position for optimal visualization. Ensure proper chamber sealing and animal orientation.

▶ 04:50
4
Acquire long-term calcium imaging data

Perform time-lapse fluorescence imaging to record neural calcium signals without immobilizing the animal. Maintain appropriate imaging intervals and duration.

▶ 05:37
5
Analyze long-term calcium imaging results

Process acquired imaging data to extract neural activity patterns and behavioral correlates across extended observation periods.

▶ 06:27
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