I have talked to a coupled of people from my institution and it seems there is no consensus on when a zebrafish embryo/larvae is called dead. Some people say it is when they cannot see any heart beat anymore, some people say it is when there is no touch and startle response anymore.
Of note: I am exposing the zebrafish embryos/larvae to toxins which have been associated with paralysis which excludes the startle and touch response in this case. Would you just use the heart beat in this case?
Are there any additional parameters by which death is judged or would you recommend a combination of parameters?
Heart beat is more definitive. Lack of touch and startle maybe due to hypoactivity after a drug treatment.
Heartbeat can be tricky. If you're using anesthesia (in my experience MS-222 or cold) can stop the heart but if you take away the treatment, the heartbeat "re-start" and the larvae behave normally.
I would recommend taking notes of several symptoms including touch response, heartbeat, and necrosis (especially in the yolk).
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