I am trying to make DG and PC lipid mixed liposomes using waterbath sonication. Does anyone have any advice on doing this without a detergent? I have 8:0 DG and 18:1 PC. I want to create liposomes and I got it to work one time, but have been unsuccessful at future attempts. The buffer I'm using contains Tris-HCl and MgCl2.
Hi Breanna
What kind of sonication bath you are using? is it "cleaning style" bath? If yes, it can be quite difficult to optimise the process. These baths usually are not very powerful to be used efficiently in biological applications. Be very "consistent" when working with sonication as the final efficiency depends on a combination of many "stupid" things like a water level in the bath, sample depth immersion in the bath, sample volume itself. He also aware that tubes used for sonication has a huge effect on the ultrasound transfer into samples.
I think it may be a cleaning style.. It is a Fisher Scientific CPXH Series 1.9L Ultrasonic bath (http://www.fishersci.com/shop/products/fisher-scientific-cpxh-series-digital-ultrasonic-cleaners-5/15337402). It does have a heater that can be used and a sonic and degas mode. The tubes I am using are Fisherbrand glass tubes (16x100mm).
Hi Breanna,
Yes, this is a cleaning bath. As I mentioned previously, they are not very powerful and it can takes a while to find out the optimal settings. Be aware that the ultrasound distribution in the bath is not equal, there are active and dead zones in the bath. SO, you can imagine that you were very lucky once when you placed your sample in an active zone. It seems that tubes you are using are flat-bottom. Usually it leads to nearly complete ultrasound reflection from a tube surface meaning that only a very limited energy was transmitted to a sample. If you are looking for more stable and adaptable solution, I could recommend the Bioruptor which is also a water bath sonicator but especially developed for biological applications. This system is used with special tubes holders and tubes allowing an optimal positioning of samples in the sonication bath and it is a way more powerful comparing to traditional cleaning baths.
https://www.diagenode.com/p/bioruptor-plus-sonication-device
Few publications using Br for lyposomes:
http://pubs.rsc.org/en/content/articlelanding/2014/ra/c4ra01334d#!divAbstract
https://www.dovepress.com/nanolipodendrosome-loaded-glatiramer-acetate-and-myogenic-differentiat-peer-reviewed-article-IJN
http://www.biochemj.org/content/401/2/597
http://www.sciencedirect.com/science/article/pii/S0196978109002307
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