I have tried ethanol drying and simple air drying (room temperature) but am unable to get a proper SEM image. I am getting structures like crystal forms, not in circular shape.
Is there any ways to get it done other than environmental SEM? My institute has only an SEM facility. Thanks in advance for your inputs and valuable time.
The liposomes dry out if you put them in vacuum, so you are imaging crystals of the buffer you prepared the liposomes in. Your could try freeze fracture to prepare the samples, but you may need a cryo SEM for this. You may be better served using TEM to image liposomes (cryo TEM may also be better for this).
Johanna, are you sure that which I had seen are crystals of buffer not liposome. Because, same pattern I see when sample gets dried out while observing under normal bright field microscope.
No facility of cryo-SEM, TEM Johanna. Should try something come out of box using SEM.
I did EDX on similar crystalline deposits from liposomes in SEM that were formed in PBS. The elemental analysis showed that the crystals were made of Na, Cl and P amongst other things i.e. the constiuents of PBS. In a liposome, the lipid bilayer forms a very small part of the whole structure, so it is unlikely that crystals of lipid will form when they dry. There might be a small amount of lipid on the crystals though. As I said above, liposomes do not survive in vacuum, they pop and dry out. Can you see your vesicles in suspension in solution under an optical or fluorescence microscope?
Johanna, Thank yous so much for your inputs. Yeah Johanna I could see very small vesicles when I focus under 40X and also in oil immersion with very high Brownian motion. Problem I am facing is with SEM analysis.
Shann, I do obviously sputter coat my sample. But problem is I am not getting the shape which I required. that is circular. May be that is not at anll my sample according to Johanna.
i think you can dry your liposomes on a Aluminum Foil in the air and then coat it with sputter
Jagathesh, thank you so much. Yeah I have seen many articles using Cryo-SEM, Env-SEM, Cryo-TEM, TEM analysis for getting the images. But problem is we dont have those facility. Only SEM that too without EDX which is becoming hell difficult to get and confirm images.
Adel, Pictures I got were not circular in shape, few are like exactly crystals.
Dear, @Adel Marzban
Can you explain for me how can i dry liposomes on a Aluminum Foil before coating it for imaging with S.E.M?
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