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I am interested in using cygel (ab109204) for medium term (up to 20 hours) live imaging of insect embryos , and possibly as a way to immobilize them for injection. I saw that there have been sucessful uses of cygel for Drosophila, but only for shorter term experiments. I saw a general recommendation to use the "sustain" variant (ab109205) for longer experiments, but this seems only related to the ability to add special media required by cells for longer term viability. Since my embryos should not interact with the medium, I guess the normal Cygel should be sufficient? Or is there something else about the "sustain" variant that makes it more suitable for long term studies?
Asked on Nov 26 2012
Thank you for contacting us.
As suggested, there is no particular reason to use CyGEL Sustain over CyGEL (a PBS-based formulation) other than the ease of adding specific media.
We have limited data on the long-term imaging of embryos other than Danio which have been imaged for 24h.
The trick is to use a cover-glass bottomed petri-dish. Make a well with an 8-10 mm diameter silicone o-ring, smeared with silicone (aka vacuum) grease, to bed it down onto the cover glass surface. Surround the o-ring well with LMP agarose – this acts as a hydration buffer to stop the hydrogel from drying out. Fill the inner well with CyGEL and your embryo(s). Lid the petri-dish. You can now image through the cover glass.
For short-term use, i.e. injections, the embryos can be immobilized in a simple shallow well and the CyGEL melted and re-set as required to best position the organism for the injection. The organism(s) can then be gently recovered by melting by re-cooling.
The lab has done quite a bit of work with Drosophila embryos of a variety of ages and they seemed OK for around an hour at least – the limit of time worked on them. They are also aware of a user in Australia has imaged an embryo for close to 2h through 5-6 cell divisions.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
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Answered on Nov 26 2012