[OpenSPIM] survival FEP Tubes

[Michael Weber]

Hey Fredrik,

thanks for watching our JoVE publication :). How old are the larvae you're trying to image? In our hands, FEP works great up to around 3 dpf. With older larvae we also see oxygen deficiency, because the tubes are gas-proof. Currently, those developmental stages are not so interesting for us, so we did not spend much time on optimizing it. We have ideas though, e.g. putting holes in the tube to allow for oxygen diffusion.

Best,
Michael


On Jan 27, 2016, at 2:51 PM, Fredrik Ek <fredrik.ek at med.lu.se> wrote:

> Hi all,
>  
> we have experienced some problems with survival of the larvae when mounted in the FEP tubes. They seems to suffocate after 1-2 h in the tube. Using the same agarose solution but keeping the larvae covered on a glass slide they stay alive and healthy. We are following the protocol for long time imaging from JOVE (Weber et al 2014) but has also tested 0.5% and 1% low melting agarose solutions for embedding. It would be much appreciated to get some input how to solve this issue.
>  
> Thanks!
>  
> Fredrik
>  
> Fredrik Ek, PhD, 
> Researcher, Chemical Biology & Therapeutics
> Department of Experimental Medical Science 
> Lund University, BMC D10 
> S-221 84 Lund, SWEDEN 
> ------------------------ 
> Fredrik.Ek at med.lu.se 
> Phone: +46-727 17 22 05 
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> OpenSPIM at openspim.org
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_____________

Michael Weber
Postdoc, Huisken lab
Max Planck Institute of Molecular Cell Biology and Genetics
Pfotenhauerstrasse 108, 01307 Dresden
Tel. 0049 351/2102837

http://www.mpi-cbg.de/huisken

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