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Hi Shalin,
Like Guy said we have done this at the Monash Live Cell Course. Normally we have used younger embryos but for older ones we have had good luck with CyGel. In the past i have done other imaging with older (96hpf) embryos and have done it just with low melt temp agarose. Its good for about 24 hours of imaging. If you need more then you have to chop away part around the tail end to allow the embryo to grow.
I also know of a group here in Melbourne that are building lab on a chip type microfuidic devices to hold embryos (2-30hpf) in place. They are really amazing little things and let you load up a heap of embroys with each one locking into place using micro fluidics. These can then be imaged without a problem
Drop me a message off list and i can put you in touch with them.
Cheers
Cam
Cameron J. Nowell
Research Facilities Manager
Monash Institute of Pharmaceutical Sciences
Monash University
399 Royal Parade
(Mail address: 381 Royal Parade)
Parkville, VIC, 3052
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________________________________________
From: Confocal Microscopy List [[log in to unmask]] on behalf of Guy Cox [[log in to unmask]]
Sent: Thursday, June 05, 2014 12:16 PM
To: [log in to unmask]
Subject: Re: Keeping zebrafish embryos in the field of view but letting them grow.
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.
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