Research Area - Nanotechnology

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Quantification and localization of carbon-based nanomaterials using the ImageStream Imaging Flow Cytometer, Contributed by Florence Gazeau, Iris Marangon and Nicole Boggetto, Université Paris Diderot / CNRS

Merck:/Freestyle/BI-Bioscience/Cell-Analysis/amnis/Amnis-Research-Images/nanotechnology.jpgThe field of nanotechnology is broad and includes studies performed in cells with materials from 1 to 100 nanometers. Many different types of experiments are being perfomed in cells with nanomaterials such as the uptake and intracellular fate of nanoparticle-drug delivery vehicles, or antibody functionalized capsules to name a few. The Imagestream®x is uniquely configured to detect and localize these small particles in cells.

Label-Free Detection and Quantification of Intracellular Carbon Nanotubes

Marangon I,et al.Nano Lett. 2012 Sep 12;12(9):4830-7. Epub 2012 Sep 4. Figure 2 Imaging flow cytometry analysis of CNT uptake by EC. (a) Examples of images captured by ImageStream®x on the bright-field, FITC, and dark-field channels and their overlays for EC labeled with 50 μg/mL of CNTs. Black spots on bright-field images show colocalization with the bright spots on dark-field images. (b) Mean signal on fluorescence (FITC) and dark-field images and mean pixel signal on bright-field image analyzed for 7000 cells show a significant increase with the CNT labeling concentration. Results are mean ± standard error of the mean (s.e.m) normalized to that of control nonlabeled cells, representative of three independent experiments (n = 3). (c) Dark-field intensity correlates with the bright-field mean pixel signal to quantify CNTs in cells. Values from 7000 cells are plotted with increasingly dark blue for increasing CNT labeling concentrations. Gray dots correspond to nonlabeled cells. Pearson correlation coefficient r = −0.68 for EC labeled with 50 μg/mL. (d) Percentage of cell population that are positive for both their dark-field intensity (DF+) and bright-field mean pixel signal (BF+) compared to the gate of control cells in the bivariate dot plot (c) and thus considered as CNT-labeled cells. Percentage are mean ± s.e.m. (n = 3).