hello, i am writing to get advice on zebrafish embryo growth.
after mating the adult zebrafish, i am collect the egg and check rate of fertilization. then, put them in an incubator for 6 hours at 28 degree. most of the embryos normally develop after 6 hours of correction. however, when i check the same zebrafish embryo in 18 hours and 24 hours after collection, 50% or more of the zebrafish embryo will stop growing and the development stage will be slightly different, what is a problem?? is it a disease problem? or is water management a problem?
if you know about this problem, please help me.
thank you.
Hi!
Are the embryos alive or dead/dying? If they are alive and growing in a slightly different rate, I would say you do not need to worry too much.
If the embryos are dead or dying, then probably it is the problem of the mating parents (I assume you didn't manipulate the embryos). You can repeat with different mating pairs, or even the same pairs after giving them a week or two rest.
Gastrulation happens a few hours after fertilisation. if the quality of the embryos are bad then this is most likely when they will die.
From my experience it is not uncommon that occasionally there are some bad clutch of embryos. If it doesn't happen too often then it should be ok.
It is not unusual to have, from some lines of parents, 30-50% of the clutch die during early stages. If the lines are important to you, keep breeding them and then propagate the ones that survive.
However, it is also possible there is some bacterial contamination in the cultures. We use these steps to prevent / minimize any problems:
- Collect eggs in strainers stored in ethanol; rinse in egg water before use.
- Autoclave the egg water that you use for incubation in your lab.
- Transfer eggs through several washings of sterile egg water before you incubate.
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