Quantifying the level of nanoparticle uptake in mammalian cells using flow cytometry

Nanoscale
HyeRim ShinJi Youn Lee

Abstract

Reliable quantification of nanoparticle uptake in mammalian cells is essential to study the effects of nanoparticles in the fields of medicine and environmental science. Most conventional quantification methods, such as electron microscopy or confocal imaging, are laborious and semi-quantitative and therefore not readily applicable to routine analyses. Here, we developed assays to quantify fluorescently labelled nanoparticle uptake in mammalian cells using a flow cytometer. The first approach was to measure the percentage of nanoparticle-containing cells based on a cutoff fluorescence intensity as set from a histogram of control cells, which is a quick and easy way to relatively compare nanoparticle uptake in the same set of experiments. The second approach was to measure the calibrated fluorescence intensity of the nanoparticle-treated cells in molecules of equivalent soluble fluorophore (MESF) values using calibration beads, which allows for comparisons between different sets of experiments. We successfully applied the developed assays to more readily measure fluorescence-labelled silica nanoparticle uptake in A549 lung carcinoma cells in a quantitative rather than semi-quantitative manner. We further tested the assays with n...Continue Reading

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Citations

Jan 20, 2021·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Daniela S ReisLidia M Andrade
Aug 5, 2021·The Science of the Total Environment·Monica Torres-RuizAna Isabel Cañas Portilla
Sep 22, 2021·Chemical Reviews·Vivian M LechnerMatthew J Gaunt
Nov 2, 2021·Environmental Science & Technology·Xiaolin ZhangLi Mu

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Methods Mentioned

BETA
transmission
X-ray
confocal microscopy
confocal
flow cytometry
dynamic light scattering
scanning electron microscopy
PMA

Software Mentioned

FlowJo
ImageJ
Excel

Related Concepts

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