Optimized CUBIC protocol for 3D imaging of chicken embryos at single-cell resolution [RESEARCH REPORT]

Maria Victoria Gomez-Gaviro, Evan Balaban, Diana Bocancea, Maria Teresa Lorrio, Maria Pompeiano, Manuel Desco, Jorge Ripoll, and Juan Jose Vaquero

The CUBIC tissue clearing protocol has been optimized to produce translucent immunostained whole chicken embryos and embryo brains. When combined with multispectral light sheet microscopy, the validated protocol presented here provides a rapid, inexpensive and reliable method for acquiring accurate histological images that preserve three-dimensional structural relationships with single-cell-level resolution in whole early-stage chicken embryos, and in the whole brains of late-stage embryos.

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