I'm currently trying to live-stain the zebrafish embryo membranes during gastrulation, ideally beginning at 3 hpf until 6 hpf. However, I can't seem to find a good stain for the task. I have tried using FM 4-64, which doesn't stain the membranes very well during this stage. Are there any other alternatives?
Hello, I will recommend fluorescent dyes PY3304, PY3174 and PY3184, whose fluorescence properties are sensitive to membrane lipid order. In artificial bilayers, the fluorescence emission spectra are red-shifted between the liquid-ordered and liquid-disordered phases.
Karen A. Darbinyan
Thanks Karen,
Do you have any suggestions as how to use these dyes, whether by soaking or directly injecting them into the embryo?
Your welcome !
soaking
Take three day-old zebrafish embryos and incubate with dyes PY3304, PY3174 or PY3184 (2 µM −5 ) dissolved in E3 media for 2 h. The embryos can be anesthetized with 50 µL of 0.04% (wt/vol) tricaine.
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