Hello,
I'm trying to see the cytotoxic effect of curcumin loaded nanoparticles (Iron Oxide) in A549 cell line but the data is absurd.
1. The optical density of wells loaded with nanoparticles alone is coming out higher than the control cells containing media only. It means the nanoparticles are scattering light at the measurement wavelength (540 or 570nm). Should I wash the wells with PBS to remove the nanoparticles, the washing step would remove some of the cells too!
2. The nanoparticles are aggregating after their addition into the 96 well plate. The aggregation is due to the salts in the media, does it affect the results too?
3. We are solubilizing curcumin in ethanol before loading into the wells, should we subtract the optical density of the wells containing drug in ethanol from wells containing ethanol only ? Also, upon increasing the concentration of curcumin I'm not able to get any trend, the optical density is coming out to be random.
Anyone has experience in doing MTT assay of nanoparticles loaded drug?
1. The optical density of wells loaded with nanoparticles alone is coming out higher than the control cells containing media only. - This is a common problem with nano-particles aggregation causing light scattering. How about using a fluorescence indicator instead? Just make sure in advance your nano-particles don't have self fluorescence at the same wavelength.
2. Aggregation could lower dramatically the bio-availability of the nano-particles to the cells.
Yoram
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