I prepared my liposome using the modified hydration-rehydration technique and used it for encapsulating silver nanoparticle. Zeta analysis gave -25mV which is quite stable but when cells (THP1) are treated with the liposome and used for flow cytometry I get many debris and some signals which i believe are artifacts due to the liposome (clearly seen in the cell suspension) in the channel. How can I ensure the liposome is completely gone before the flow analysis. it is disturbing the results.
Hi Azeez, I am also working with THP1 cells. What I did was I collected the cells (suspension) and centrifuge the them down at low speed (300g x 5mins). At this speed, your liposomes are still floating in the supernatant. Washing the cell 2 times should be enough to get rid of the liposomes.
The debris can also come from dead cells, which may affect your result in FACS. So you can use viability dye such as DAPI, eflor780, or Zombie to stain live-dead cells and gating only for the live cells in you FACS. These viability dye won't stain your liposome as it stains only for dead cells.
The liposome are not as stable as you are guessing! The cleaning of the preparation is another requirement which can be accomplished through centrifugation. The clearance of your preparation from the unwanted and dead cells is another factor which has rightly been suggested by Ms Kanokwan, needs to be implemented in full.
@Kanokwan Sansanaphongpricha Thanks a lot for your idea. I will definitely try that and see how it goes.
@Riaz A. Khan. Yes thats true. I will try that and I expect it to work out fine.
Thanks a lot for your contributions.
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