PUBLICATION
In vivo Imaging of Calcium Activities from Pancreatic β-cells in Zebrafish Embryos Using Spinning-disc Confocal and Two-photon Light-sheet Microscopy
- Authors
- Zhao, J., Liang, S., Zhao, Y., Zong, W., Tran, E., Chen, L., Liu, Y.
- ID
- ZDB-PUB-220111-12
- Date
- 2021
- Source
- Bio-protocol 11: e4245 (Journal)
- Registered Authors
- Chen, Liangyi, Liu, Yanmei
- Keywords
- Calcium imaging, In vivo imaging, Pancreatic β-cells, Spinning-disc confocal microscopy, Two-photon light-sheet microscopy, Zebrafish embryo, β-cell function
- MeSH Terms
- none
- PubMed
- 35005090 Full text @ Bio Protoc
Citation
Zhao, J., Liang, S., Zhao, Y., Zong, W., Tran, E., Chen, L., Liu, Y. (2021) In vivo Imaging of Calcium Activities from Pancreatic β-cells in Zebrafish Embryos Using Spinning-disc Confocal and Two-photon Light-sheet Microscopy. Bio-protocol. 11:e4245.
Abstract
Visualizing the function of pancreatic β-cells in vivo has been a long-sought goal for β-cell researchers. Unlike imaging of β-cells in mammalian species with conventional positron emission tomography and single-photon emission computed tomography, which only provides limited spatial-temporal resolution, transparent zebrafish embryos are a unique model that allows high-resolution fluorescent imaging of β-cells in their native physiological microenvironment in vivo. Here, we detail a protocol for real-time visualization of individual β-cell function in vivo in a non-invasive manner, through combination of a novel transgenic zebrafish reporter line Tg (ins:Rcamp1.07) with both a commercial spinning-disc confocal microscope and an in-house developed super-resolution microscope (2P3A-DSLM). The protocol described here allows for the longitudinal monitoring of dynamic calcium activities from heterogeneous β-cells in early developing zebrafish embryos and is readily adaptable for use in imaging other important processes in islet biology, as well as screening new compounds that can promote β-cell function or maturation using a living whole organism system.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping