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A Novel Flow‐Cytometry‐Based Assay for Cellular Uptake Studies of Polyelectrolyte Microcapsules
Author(s) -
Semmling Maximilian,
Kreft Oliver,
Muñoz Javier Almudena,
Sukhorukov Gleb B.,
Käs Josef,
Parak Wolfgang J.
Publication year - 2008
Publication title -
small
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.785
H-Index - 236
eISSN - 1613-6829
pISSN - 1613-6810
DOI - 10.1002/smll.200800596
Subject(s) - flow cytometry , fluorescence , polyelectrolyte , biophysics , chemistry , cytometry , endosome , membrane , chromatography , cell , biochemistry , microbiology and biotechnology , biology , polymer , physics , organic chemistry , quantum mechanics
A flow‐cytometry‐based assay is presented with which the uptake of polyelectrolyte capsules can be quantified. The cavity of the capsules is loaded with the pH‐sensitive dye SNARF, which emits in the red and green in alkaline and acidic environments, respectively. By recording the fluorescence intensities in the red and green channels, the localization of capsules associated with cells can be determined. Capsules adherent to the outer cell membrane fluoresce in the red due to the alkaline pH of the cell medium, whereas capsules internalized by cells fluoresce in the green due to the acidic pH in the endosomal/lysosomal/phagosomal compartments in which incorporated capsules are located. Adding the SNARF readout to the scattering signal typically derived with flow cytometry analysis allows for a more detailed quantitative analysis of particle uptake, which can also distinguish between adherent and ingested particles.