Protocol detail

Microscopic Imaging of C. elegans Embryos

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For ex utero imaging, embryos or oocytes were dissected from hermaphrodite worms into Shelton’s Growth Medium [64 (link)], supplemented with 20 μm polystyrene beads to act as spacers between glass slide and coverslip. For in utero imaging, whole worms were mounted between a 10% M9 agarose pad and coverslip, in M9 containing either 0.1 μm polystyrene beads (Polysciences), or 5% tetramisole in order to immobilize the worms.

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Reich J.D., Hubatsch L., Illukkumbura R., Peglion F., Bland T., Hirani N, & Goehring N.W. (2019). Regulated Activation of the PAR Polarity Network Ensures a Timely and Specific Response to Spatial Cues. Current Biology, 29(12), 1911-1923.e5.

Publication 2019

polystyrene beads

Agarose Embryos Growth medium Hermaphrodite Immobilize Oocytes Polystyrene Tetramisole Utero Worms

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Variable analysis

independent variables

Ex utero imaging: presence of 20 μm polystyrene beads
In utero imaging: presence of 0.1 μm polystyrene beads or 5% tetramisole

dependent variables

Imaging of embryos or oocytes

control variables

Shelton's Growth Medium for ex utero imaging
M9 medium for in utero imaging
Mounting techniques (glass slide and coverslip for ex utero, agarose pad and coverslip for in utero)

controls

Positive control: Not specified
Negative control: Not specified

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