My question is if we load a fluorescent compound as a model drug, Is it the right way to analyze the nanoparticle by flow cytometry?
Hi, Rashmirekha.
Fluorescence activated cell sorter (FACS). As the machine's name indicated, it is specified to separate cells of the eukaryotic size range of ~10 to 30um in diameter. Nano- to a few micropartices are probably too small (well below the minimal size and fluorescence intensity to be measured).
Another issue is that distances among the nanoparticles flown are extremely small, it hard for FACS machine to differentiate whether two or more nanoparticles are separated or they are within a diameter range of a cell. IMPOSSIBLE!!!
However, if their numbers are enough to show above the threshold fluorescence of a specific cell which contains endocytosed or engulfed nanoparticles, che cell fractionation is possible. Which is not your intention.
As pointed by Dr Lee, nano size range would be too small and might not be the right technique.
If the drug has aqueous solubility, you can dialyze the nanoparticles or pass them through a gel filtration column to remove any unincorporated drug. Then you can dissolve the nanoparticles in a solvent and measure the concentration of fluorescent drug, either by fluorescence or absorbance.
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