Visualizing Neural Crest Cell Migration in a Zebrafish Embryo Using Multi-Photon Time Lapse Imaging

Abstract

Source: Williams, A. L., et al. Multi-Photon Time Lapse Imaging to Visualize Development in Real-time: Visualization of Migrating Neural Crest Cells in Zebrafish Embryos. J. Vis. Exp. (2017)

This video showcases a multi-photon time-lapse imaging protocol to visualize the migration of neural crest cells in a zebrafish embryo in real-time with high resolution.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Microscope Set-up for Time-lapse Imaging

1. Determining laser settings

1. Determining laser wavelength to use. Use a wavelength that is twice the excitation wavelength of the fluorophore of interest (e.g. wavelength between 880 and 940 nm for green fluorescent protein (GFP).
NOTE: In the present study, the wavelength setting for GFP was between 880 and 940 nm. The higher the wavelength, the lower the ou....

Results

Figure 1. Setup. A. Each component of the embryo mounting apparatus, including the.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
TC SP5 MP Multi-photon microscope	Leica Microsystems CMS GmbH
Mai Tai DeepSee Ti-Sapphire Laser	SpectraPhysics
Laser Safety Box	Leica Microsystems CMS GmbH
Leica Application Suite X (LAS X) Software	Leica Microsystems CMS GmbH
Photoshop CS 6 Version 13.0 x64 Software	Adobe
iMovie Version 10.1.4 Software	Apple
Open Bath Chamber	Warner Instruments	RC-40HP	High Profile
Quick Exchange Platform	Warner Instruments	QE-1	35 mm
Stage Adapter	Warner Instruments	SA-20LZ-AL	16.5 x 10 cm
Low-Melt Agarose	ISC Bioexpress	E-3112-25	GeneMate Sieve GQA Low MeltAgarose, 25 g
Glass Coverslips	Fisher Scientific	12-545-102	Circle cover glass. 25 mm diameter
High Vacuum Grease	Fisher Scientific	14-635-5C	2.0-lb. tube. DOW CORNINGCORPORATION1658832