Xenopus Oocyte Imaging Workflow

Xenopus laevis oocytes (Dumont stage V or VI) were purchased (Ecocyte Bioscience US LLC, Austin, Texas) and handled as described (Maksaev & Haswell, 2015 (link)). Oocytes were injected with 50 nl of 1000 ng/μl of RNA the day after isolation. Fluorescent imaging of the oocytes was carried out 48–72 h after injection. Oocytes were mounted on concave slides and covered with coverslips. Confocal imaging of the periphery of the oocytes was performed using Olympus Fluoview 1000 with BX61 microscope and the Olympus FV10-ASW software suite.

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Maksaev G., Shoots J.M., Ohri S, & Haswell E.S. (2018). Nonpolar residues in the presumptive pore‐lining helix of mechanosensitive channel MSL10 influence channel behavior and establish a nonconducting function. Plant Direct, 2(6), e00059.

Publication 2018

Fluoview 1000 with BX61 microscope FV10-ASW software suite

Austin Isolation Microscope Oocytes Xenopus laevis

Corresponding Organization : Washington University in St. Louis

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Variable analysis

independent variables

RNA concentration (1000 ng/μl)

dependent variables

Fluorescent imaging of the oocytes

control variables

Xenopus laevis oocytes (Dumont stage V or VI)
Oocyte handling as described in Maksaev & Haswell, 2015
Oocyte injection volume (50 nl)
Imaging timeframe (48-72 h after injection)
Imaging setup (Olympus Fluoview 1000 with BX61 microscope and Olympus FV10-ASW software)

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