PUBLICATION
Full-field exposure of larval zebrafish to narrow waveband LED light sources at defined power and energy for optogenetic applications
- Authors
- Burton, A.H., Jiao, B., Bai, Q., Van Laar, V.S., Wheeler, T.B., Watkins, S.C., Bruchez, M.P., Burton, E.A.
- ID
- ZDB-PUB-231102-6
- Date
- 2023
- Source
- Journal of Neuroscience Methods 401: 110001 (Journal)
- Registered Authors
- Burton, Edward A.
- Keywords
- FAP, GAVPO, Kaede, LED, MG2I, Singlet oxygen, dL5**, energy, optogenetics, power, zebrafish
- MeSH Terms
-
- Animals
- Larva
- Neurons/physiology
- Optogenetics*/methods
- Transgenes
- Zebrafish*/physiology
- PubMed
- 37914002 Full text @ J. Neurosci. Methods
Citation
Burton, A.H., Jiao, B., Bai, Q., Van Laar, V.S., Wheeler, T.B., Watkins, S.C., Bruchez, M.P., Burton, E.A. (2023) Full-field exposure of larval zebrafish to narrow waveband LED light sources at defined power and energy for optogenetic applications. Journal of Neuroscience Methods. 401:110001.
Abstract
Background Optogenetic approaches in transparent zebrafish models have provided numerous insights into vertebrate neurobiology. The purpose of this study was to develop methods to activate light-sensitive transgene products simultaneously throughout an entire larval zebrafish.
New method We developed a LED illumination stand and microcontroller unit to expose zebrafish larvae reproducibly to full field illumination at defined wavelength, power, and energy.
Results The LED stand generated a sufficiently flat illumination field to expose multiple larval zebrafish to high power light stimuli uniformly, while avoiding sample bath warming. The controller unit allowed precise automated delivery of predetermined amounts of light energy at calibrated power. We demonstrated the utility of the approach by driving photoconversion of Kaede (398nm), photodimerization of GAVPO (450nm), and photoactivation of dL5**/MG2I (661nm) in neurons throughout the CNS of larval zebrafish. Observed outcomes were influenced by both total light energy and its rate of delivery, highlighting the importance of controlling these variables to obtain reproducible results.
Comparison with existing methods Our approach employs inexpensive LED chip arrays to deliver narrow-waveband light with a sufficiently flat illumination field to span multiple larval zebrafish simultaneously. Calibration of light power and energy are built into the workflow.
Conclusions The LED illuminator and controller can be constructed from widely available materials using the drawings, instructions, and software provided. This approach will be useful for multiple optogenetic applications in zebrafish and other models.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping