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Absolute Quantitation of Sub‐ M icrometer Particles in Cells by Flow Cytometry
Author(s) -
Höcherl Anita,
Landfester Katharina,
Mailänder Volker
Publication year - 2013
Publication title -
macromolecular bioscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.924
H-Index - 105
eISSN - 1616-5195
pISSN - 1616-5187
DOI - 10.1002/mabi.201300182
Subject(s) - flow cytometry , fluorescence , micrometer , chemistry , confocal laser scanning microscopy , nanoparticle , particle (ecology) , confocal , fluorescence microscope , confocal microscopy , analytical chemistry (journal) , quantitative analysis (chemistry) , cytometry , nanotechnology , microscopy , hela , particle size , biophysics , materials science , chromatography , in vitro , microbiology and biotechnology , biochemistry , optics , oceanography , physics , geology , biology
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 μL 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.
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