I am conducting the uptake study of different nanoparticle formulations on splenocytes. Their cell uptake was then assessed by comparing MFI data from flow cytometry. To do this, we did labeling nanoparticles using the fluorescent dye Dylight633. The number of Dylight633 molecules per 1 mol of nanoparticle may be different for each formulation. Can we compare the cell uptake property of nanoparticle formulations by directly comparing their MFI values or we need to do normalisation based on the number of Dylight633 molecules per 1 mol of nanoparticle?
If you have two batches of NP, one bright and one dim, then the MFI for splenocytes that took up the bright NP will be higher than the splenocytes that took up the dim NP when the number of particles taken up is equal. So you can't directly compare MFI to tell which NP is taken up more. Ideally you would normalize the labeling, so that all NPs get the same number of Dylight633 per mol of NP. Normalization by dividing MFI by the number of Dylight633 per mol seems like an over-simplification and I would be worried that it is not safe to assume that doubling the number of dylight molecules should double MFI. Because of this issue, I don't think that this method provides a good basis for comparison of uptake between different types of particles, but can be used for comparing uptake of the same particle at different conditions.
- Badges
- Science method