Protocol for simultaneous in vivo two-photon imaging and locomotion quantification during olfactory stimulation and pharmacology in walking Drosophila
Abstract
To decipher the circuit and molecular mechanisms underlying neural information processing, it is essential to manipulate cellular targets while simultaneously recording neural activity and behavior during sensory stimulation. Here, we present a protocol for Drosophila that temporally synchronizes olfactory stimulation and pharmacological intervention with in vivo recording as well as locomotion measurement. We detail step-by-step workflows for head-fixed walking flies, including fly preparation, surgery, two-photon imaging, and locomotion detection, providing instructions for surgery station, odor stimulator, and perfusion system setup.
Fields of Study & Subjects
Neurobiology and Insect Physiology Research
Zebrafish Biomedical Research Applications
Nerve injury and regeneration
Publication details
- Journal / source
- STAR Protocols
- Publisher
- Elsevier BV
- Volume
- 7
- Issue
- 3
- Pages
- 104659–104659
- Publication date
- 25 June 2026
- Language
- EN