Absolute Quantitation of Sub-Micrometer Particles in Cells by Flow Cytometry

作者：Hoecherl Anita; Landfester Katharina; Mailaender Volker

来源：Macromolecular Bioscience, 2013, 13(11): 1568-1575.

DOI：10.1002/mabi.201300182

• absolut quantitation
• confocal laser scanning microscopy
• flow cytometry
• intracellular uptake
• nanoparticles

Absolute quantitative measurements of nanoparticle (NP) uptake are a prerequisite to determine doses of NPs in pharmacological and toxicological studies. However, absolute quantitation is rarely reported, hindering the comparison between different studies. Here, a new flow cytometric approach is presented to analyze fluorescent NPs with a standard non-scanning flow cytometer and to quantify them inside cells. The mean fluorescence intensity of a single particle and the particle concentration (NPs per L medium) are obtained. A routine for rapid quantitative counting of the endocytosed NPs in HeLa cells by flow cytometry (FC) is developed and validated by confocal laser scanning microscopy. As a proof-of-concept, the quantitative measurements show that the cellular uptake efficiency of negatively charged poly(methyl methacrylate) NPs is very low, that is, in the range of 10(-3)% of the added particle amount.

• 出版日期2013-11