Embryo Fixation and Serial Block-Face SEM

Embryos were fixed in 2.5% glutaraldehyde in embryo grade water (Sigma, W1503) for 1–2 h at room temperature. Fixed embryos were rinsed with 1x PBS and stored at 4 °C before processing. Processing was performed exactly as described previously^{76 (link)}. Serial blockface sectioning and scanning electron microscopy were performed using a VolumeScope serial block-face EM (Thermo Fisher, Waltham, MA) equipped with a low-vacuum backscatter detector (VS-DBS; Thermo Fisher).

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Hawdon A., Geoghegan N.D., Mohenska M., Elsenhans A., Ferguson C., Polo J.M., Parton R.G, & Zenker J. (2023). Apicobasal RNA asymmetries regulate cell fate in the early mouse embryo. Nature Communications, 14, 2909.

Publication 2023

W1503 VolumeScope serial block-face EM VS-DBS

Embryos Face Glutaraldehyde Scanning electron microscopy Vacuum

Corresponding Organization :

Other organizations : Australian Regenerative Medicine Institute, Monash University, Walter and Eliza Hall Institute of Medical Research, University of Melbourne, University of Adelaide, Centre for Cancer Biology, University of Duisburg-Essen, University of Queensland

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Variable analysis

independent variables

Fixation of embryos in 2.5% glutaraldehyde in embryo grade water (Sigma, W1503) for 1-2 h at room temperature

dependent variables

Visualization and examination of embryos using serial blockface sectioning and scanning electron microscopy

control variables

Rinsing of fixed embryos with 1x PBS before storage at 4°C
Processing performed exactly as described previously (link provided)

controls

No positive or negative controls were explicitly mentioned in the provided information.

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