Agar Mounting: A Basic Method of Mounting Live Zebrafish Embryos for Long-Term Imaging
Published: April 30, 2023
Abstract
Source: Hagedorn E. J., et al. Generation of Parabiotic Zebrafish Embryos by Surgical Fusion of Developing Blastulae. J. Vis. Exp. (2016).
This video describes step by step instructions on mounting live zebrafish embryos on 6 well plate using low melt agarose. It also provides the process to acquire images of live embryos and analyze it using software.
Protocol
1. Microscope Setup, Image Acquisition, Processing and Analysis Prepare 0.8% low melting point (LMP) agarose: Add 0.8 g LMP agarose to 100 ml E3 and heat until fully dissolved. While hot, aliquot 1 ml of LMP agarose into 1.5 ml microfuge tubes in a 37 °C heating block. The LMP agarose will remain molten at 37 °C. Add 40 µl of 4 mg/ml (25x), pH 7.0 tricaine to each 1 ml aliquot of LMP agarose at 37 °C. NOTE: If working with a pigmented strain, add 20 µ…
Materials
| LMP agarose (Ultrapure LMP agarose) | Invitrogen | 16520100 |
| Tricaine (powder) (Tricaine Methanesulfonato, Tricaine-S) | Western Chemical Inc | MS 222 |
| Glass-bottom 6-well plates used for imaging | MatTek | P06G1.5-20-F |
| 10 ml pipette pump (green) (Pipette Pump Pipettor) | Novatech International | F37898-0000 |
Citer Cet Article
Agar Mounting: A Basic Method of Mounting Live Zebrafish Embryos for Long-Term Imaging. J. Vis. Exp. (Pending Publication), e20187, doi: (2023).