Imaging and Tracking Embryonic Development

Embryos were anesthetized with 0.01% 3-aminobenzoic acid ester (Tricaine), immersed in 0.8% low-melting point agarose and mounted in glass-bottomed 35 mm Petri dishes (Electron Microscopy Sciences). After mounting, the Petri dish was filled with egg water containing PTU and Tricaine. A 40× water objective (NA = 1.1) mounted on a motorized Zeis AxioObserver Z1 microscope equipped with a Quorum WaveFX-X1 (Quorum Technologies) or Andor CSU-W1 (Oxford Instruments) spinning disc confocal system was used to capture all images. Images were imported into MetaMorph (Molecular Devices) and/or ImageJ. Time-lapse images were analyzed using cell tracking software as previously described in (Wang et al. 2018 (link)). All images were then imported in Adobe Photoshop and Illustrator. Adjustments were limited to levels, contrast, and cropping.

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Ali M.F., Latimer A.J., Wang Y., Hogenmiller L., Fontenas L., Isabella A.J., Moens C.B., Yu G, & Kucenas S. (2021). Met is required for oligodendrocyte progenitor cell migration in Danio rerio. G3: Genes|Genomes|Genetics, 11(10), jkab265.

Publication 2021

glass-bottomed 35 mm Petri dishes AxioObserver Z1 microscope Quorum WaveFX-X1 Andor CSU-W1 MetaMorph

Agarose Aminobenzoic acid Devices Dish Electron microscopy Embryos Ester Microscope Tricaine

Corresponding Organization : University of Virginia

Other organizations : Virginia Tech, Fred Hutch Cancer Center

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Variable analysis

independent variables

Concentration of 3-aminobenzoic acid ester (Tricaine) used to anesthetize embryos (0.01%)

dependent variables

Ability to capture images of embryos using the motorized Zeis AxioObserver Z1 microscope with a 40x water objective (NA = 1.1) and a Quorum WaveFX-X1 or Andor CSU-W1 spinning disc confocal system

control variables

Mounting of embryos in 0.8% low-melting point agarose in glass-bottomed 35 mm Petri dishes (Electron Microscopy Sciences)
Filling of Petri dish with egg water containing PTU and Tricaine after mounting embryos
Use of cell tracking software to analyze time-lapse images, as previously described in Wang et al. 2018

positive controls

None specified

negative controls

None specified

Annotations

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