time-lapse imaging of embryos

Model Organism: Worms Methods & Reagents
1

Hi everybody,

Does anyone know why my early embryos get arrested after few cell divisions when I try to do time-lapse imaging? I use the 63x objective and the embryos are mounted on 2-3% agarose with the help of some M9. The temperature of the room is set to 22C.

Thanks a lot

PS. The strain I use is carrying a prom::GFP transgene

2

I am guessing you’re using a confocal with a fluorescent bulb of some sort. Early embryos are extremely sensitive to the radiation (UV?) that comes from standard mercury lamps and the new wave (HXP, Excite) ones as well. You need to turn your power WAY down and your imaging time up. Or try a spinning disk confocal, where the scan times can be fast.

The embryos will probably croak at some point regardless but you should be able to get a few more cycles in.

Steve

3

That was it, I tuned down the power of the UV lamp and embryos are happily growing!

Thanks Steve!

K

4

Glad to hear it!