PUBLICATION
In vivo whole-brain imaging of zebrafish larvae using three-dimensional fluorescence microscopy
- Authors
- Cho, E.S., Han, S., Kim, G., Eom, M., Lee, K.H., Kim, C.H., Yoon, Y.G.
- ID
- ZDB-PUB-230516-30
- Date
- 2023
- Source
- Journal of visualized experiments : JoVE (194): (Journal)
- Registered Authors
- Eom, Minho, Kim, Cheol-Hee
- Keywords
- none
- MeSH Terms
-
- Animals
- Brain*/diagnostic imaging
- Brain*/physiology
- Imaging, Three-Dimensional/methods
- Larva/physiology
- Microscopy, Fluorescence/methods
- Neuroimaging
- Sepharose
- Zebrafish*/physiology
- PubMed
- 37184275 Full text @ J. Vis. Exp.
Citation
Cho, E.S., Han, S., Kim, G., Eom, M., Lee, K.H., Kim, C.H., Yoon, Y.G. (2023) In vivo whole-brain imaging of zebrafish larvae using three-dimensional fluorescence microscopy. Journal of visualized experiments : JoVE. (194):.
Abstract
As a vertebrate model animal, larval zebrafish are widely used in neuroscience and provide a unique opportunity to monitor whole-brain activity at the cellular resolution. Here, we provide an optimized protocol for performing whole-brain imaging of larval zebrafish using three-dimensional fluorescence microscopy, including sample preparation and immobilization, sample embedding, image acquisition, and visualization after imaging. The current protocol enables in vivo imaging of the structure and neuronal activity of a larval zebrafish brain at a cellular resolution for over 1 h using confocal microscopy and custom-designed fluorescence microscopy. The critical steps in the protocol are also discussed, including sample mounting and positioning, preventing bubble formation and dust in the agarose gel, and avoiding motion in images caused by incomplete solidification of the agarose gel and paralyzation of the fish. The protocol has been validated and confirmed in multiple settings. This protocol can be easily adapted for imaging other organs of a larval zebrafish.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping