Drosophila In Vivo Calcium Imaging: A Method for Functional Imaging of Neuronal Activity
Published: April 30, 2023
Abstract
Source: Hancock, C. E., et al. In Vivo Optical Calcium Imaging of Learning-Induced Synaptic Plasticity in Drosophila melanogaster. J. Vis. Exp. (2019).
This video describes in vivo calcium imaging in Drosophila neurons using GCaMP. The featured protocol clip shows how to record changes in GCaMP fluorescence from mushroom body neurons during an olfactory conditioning experiment.
Protocol
This protocol is an excerpt from Hancock et al., In Vivo Optical Calcium Imaging of Learning-Induced Synaptic Plasticity in Drosophila melanogaster, J. Vis. Exp. (2019). 1. In vivo calcium imaging Use a multiphoton microscope equipped with an infrared laser and a water immersion objective (see Table of Materials), installed on a vibration-isolated table. For the visualization of GFP-based calcium indicators, …
Materials
| 1-Octen-3-ol | Sigma-Aldrich, St. Louis, MO, USA | O5284 | Chemical used as odorant |
| 3-Octanol | Sigma-Aldrich, St. Louis, MO, USA | 218405 | Chemical used as odorant |
| 4-Methylcyclohexanol | Sigma-Aldrich, St. Louis, MO, USA | 153095 | Chemical used as odorant |
| Bandpass filter for EGFP (525/50 nm) | Carl Zeiss Microscopy GmbH, Jena, Germany | ||
| Mineral oil | Sigma-Aldrich, St. Louis, MO, USA | M8410 | Used as diluent for odorants |
| Mode-locked Ti-Sapphire laser Chameleon Vision 2 | Coherent Inc., Santa Clara, CA, USA | Tunable infrared femtosecond laser | |
| Multiphoton Microscope LSM 7MP equipped with BiG detectors | Carl Zeiss Microscopy GmbH, Jena, Germany | Multiphoton microscope, multiple companies provide similar devices. | |
| Plan-Apochromat 20x (NA = 1.0) water immersion objective | Carl Zeiss Microscopy GmbH, Jena, Germany | 421452-9900-000 | Objective W "Plan-Apochromat" 20x/1.0 DIC M27 70mm |
| Visual Basics of Applicatons (VBA) software to receive a trigger from the odor-delivery device and the electric shock application device (power supply) to interact with the ZEN software from Zeiss that controls the microscope | Custom-written and available upon request | n.a. | n.a. |
Diesen Artikel zitieren
Drosophila In Vivo Calcium Imaging: A Method for Functional Imaging of Neuronal Activity. J. Vis. Exp. (Pending Publication), e20129, doi: (2023).