Shalin,

                I know they do this sort of thing in the Monash Micro-Imaging Live Cell course so maybe one of the Steves will reply.  My own suggestion would to punch a hole just smaller than the FOV in your agarose, so the embryo can grow in any direction without you losing track of it.  A cow-sized hypodermic needle would probably do it.  (I've got several but I think it would be easier if you sourced them locally).  

                                Guy

-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Shalin Mehta
Sent: Thursday, 5 June 2014 6:15 AM
To: [log in to unmask]
Subject: Keeping zebrafish embryos in the field of view but letting them grow.

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Hello listers,
We are imaging zebrafish embryos with 10x and the embryos just about fit the field of view at the development stage of interest (4hpf-30hpf).

Are there tried and tested methods for giving the embryos room to grow, while keeping them within the field of view?

Embryos are mounted in agarose and we can make space in agarose pad near the tail when the embryos are older. But, identifying the growth axis at young stages (~4hpf) is difficult.

All inputs appreciated,
Thanks

Assistant Research Scientist,
Marine Biological Laboratory,
7 MBL Street, Woods Hole MA 02543, USA

website: http://mshalin.com
(office) Lillie 110, (ph) 508-289-7374.