Issue 29, 2020

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

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 nine different types of mammalian cells and investigated the correlation between cell type/size and nanoparticle uptake.

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

Supplementary files

Article information

Article type
Paper
Submitted
26 Feb 2020
Accepted
29 Jun 2020
First published
02 Jul 2020
This article is Open Access
Creative Commons BY-NC license

Nanoscale, 2020,12, 15743-15751

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

H. Shin, M. Kwak, T. G. Lee and J. Y. Lee, Nanoscale, 2020, 12, 15743 DOI: 10.1039/D0NR01627F

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