Hi everyone,
I am trying to make SLB by liposome rupture, recently i found the SLB did form, however, after bleaching the hole could not completely recover. I think there exists multi-layers.
For liposome compositions,
POPC 80%
DOPS 20%
Rhod-PE 0.5%
100 uM (total lipids) were added onto the acid-etched glass coverslip, followed by incubation at room temperature for 20 minutes, then washing out the excess liposomes.
My question is, is it multi-layers? if so, what is the optimal SUV/LUV concentration or incubation conditions for SLB formation?
Many thanks!
Hi,
In order to help you with your question, please clarify these sections of your question:
You mentioned: "... after bleaching the hole could not completely recover ..."!?
Also, there is a contradiction in your other sentence:
"My question is, is it multi-layers? if so, what is the optimal SUV/LUV concentration or incubation conditions for SLB formation?"
Note: As you know SUV and LUV are not multilayered structures. Therefore, it must be clarified that the problem you are facing in your experiment is due to the nature of the liposomes you are manufacturing (perhaps they are not SUV or LUV in the first place) , or is it because of the "bleaching" procedure you mentioned above?
Meanwhile, have a look at these recent publications relevant to your work:
Luchini, A., Nzulumike, A. N., Lind, T. K., Nylander, T., Barker, R., Arleth, L., ... & Cárdenas, M. (2019). Towards biomimics of cell membranes: Structural effect of phosphatidylinositol triphosphate (PIP3) on a lipid bilayer. Colloids and Surfaces B: Biointerfaces, 173, 202-209.
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Imai, K., Horio, T., Hattori, T., Sawada, K., & Tero, R. (2019). Lipid bilayer formation on an ion image sensor and measurement of time response of potential dependency on ion concentration. Japanese Journal of Applied Physics, 58(SD), SDDK06.
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Combet, S., Cousin, F., Rezaei, H., & Noinville, S. (2019). Membrane interaction of off-pathway prion oligomers and lipid-induced on-pathway intermediates during prion conversion: A clue for neurotoxicity. Biochimica et Biophysica Acta (BBA)-Biomembranes, 1861(2), 514-523.
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Dear Shen Wang ,
Did you came across the following excellent review?
Lind, T. K., & Cárdenas, M. (2016). Understanding the formation of supported lipid bilayers via vesicle fusion—A case that exemplifies the need for the complementary method approach. Biointerphases, 11(2), 020801.
https://avs.scitation.org/doi/full/10.1116/1.4944830
Or this more than excellent tutorial?
Kurniawan, J., Ventrici de Souza, J. F., Dang, A. T., Liu, G. Y., & Kuhl, T. L. (2018). Preparation and Characterization of Solid-Supported Lipid Bilayers Formed by Langmuir–Blodgett Deposition: A Tutorial. Langmuir, 34(51), 15622-15639.
https://pubs.acs.org/doi/abs/10.1021/acs.langmuir.8b03504
My impression is that the size of the vesicles (as small as possible) is not the only factor of importance. Matters like choice of salts, temperature (and temperature cycles) are equally or even more important.
Hope these references will help you any further.
Best regards.
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