After electroformation, our unilamellar vesicles act very dynamic and unstable on BSA passivated slides (especially as their sizes get bigger). We sometimes observe them bursting and reforming. Also most of the vesicles have tubular structures inside which we want to get rid of. The buffer use inside is glucose solution and outside we have small amounts of salt with very similar osmolarity. I would be happy to hear any suggestions. Thanks!
Dear Feyza,
may I ask how similar the osmolarity is, did you measure the solutions? That's my first idea and very important.
Best
Annika
Dear Feyza,
I would also recommend checking the inner solution and outer solution osmolarities. Large osmolarity differences between inner and outer solution osmolarity will cause these effects. Also, are you using monovalent salts or di-/tri-valent salts? Are you getting patches of bilayers on your BSA-passivated slides?
Good luck!
Suzanne
Thanks for your answers. The inner buffer osmolarity is 349 mOsm/L and the outer buffer is 341 mOsm/L. I keep the difference under 10 mOsm/L in all experiments.
Inner buffer is just sucrose solution and in the outer buffer we have Hepes and KCl.
In the BSA passivated slides we do not see patches due to rupturing. But maybe their contact still affect the stabilities somehow.
1)Make sure your BSA buffer has a matched osmolarity in case you have residual BSA buffer left.
2) What are your lipids? Some lipids may not be able to form stable vesicles.
3) How long do you electroform for? Too long can cause oxidation of lipids.
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