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Cell Biology JoVE (Open Access) Citable · DOI

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

DOI: 10.3791/51483-v
What you'll learn
  • ✓ Design and assemble laser coupling systems for optogenetic stimulation
  • ✓ Perform in vivo optogenetic stimulation in rodent CNS during behavior
  • ✓ Interpret behavioral outputs from optogenetic manipulation experiments
Protocol

Optogenetics has become a powerful tool for use in behavioral neuroscience experiments. This protocol offers a step-by-step guide to the design and set-up of laser systems, and provides a full protocol for carrying out multiple and simultaneous in vivo optogenetic stimulations compatible with most rodent behavioral testing paradigms.

Difficulty
advanced
Total time
~2–4 weeks (including surgery recovery, viral expression, and behavioral testing per animal)
Model organism
Rodent (mouse/rat)
Biosafety
BSL-1

Steps

1
Design and couple non-contact laser system

Assemble laser delivery optics and coupling apparatus for optogenetic stimulation without direct fiber contact. Align collimation and focus optics to maximize light delivery efficiency to implanted brain regions.

▶ 01:46
2
Perform in vivo optogenetic stimulation surgery

Conduct surgical implantation of optical fibers or cranial windows, establish stimulation parameters, and deliver light pulses to targeted CNS regions during the experiment. Monitor post-operative recovery and electrode/optical path integrity.

▶ 05:13
3
Quantify rodent behavioral responses to stimulation

Record and analyze behavioral outputs (locomotion, freezing, approach/avoidance) evoked by optogenetic manipulation. Compare responses across stimulation conditions and control trials.

▶ 08:00
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