Hi,
When making liposomes, the lipids in organic solvent are first put in a round-bottom flask and are 'rotated' while getting dried under nitrogen gas to obtain "a uniform thin film" of lipids on the walls of the round bottom flask. And then the lipids are hydrated, destabilized with detergent, etc.
What is the reason of trying to get a thin film of dried lipid by rotating the flask?
Why can't we just dry the lipids in the flask while the flask is just standing straight up and get a thick layer of lipids at the bottom and then hydrate the lipids?
Many thanks for your help in advance!
Dear Yn So. Always good to question why ‘we’ do things the way we are used to do them. I can think of a number of reasons why the way you described it is a good practice:
-The need to get rid of the organic solvent. Lipids are first dissolved in organic solvent and in most cases, you don’t want to end up with these type of solvents in your actual experiment (toxic, can disturb the experiment etc.).
-The best way to get rid of the organic solvent. In order to get rid of the organic solvent (even trace amounts) it is better to create a thin film (so the surface of evaporation is maximized)
-The best way to (re)solubilise the lipids. Additionally, the thin film ensures the creation of good solubilization of the lipids in the final (buffer) solution (some lipids like anionic phospholipids will solubilize in aqueous solutions faster and when the film is thick then you might get microsomes/liposomes where the various lipids are not uniformly distributed)
Additionally, I think it is always a good thing that the preparation of samples occurs in a well-described and reproducible way.
Best regards.
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