A fast analysis method to quantify nanoparticle uptake on a single cell level

Author:

Torrano Adriano A1,Blechinger Julia1,Osseforth Christian1,Argyo Christian1,Reller Armin2,Bein Thomas1,Michaelis Jens13,Bräuchle Christoph4

Affiliation:

1. Ludwig-Maximilians-University Munich, Department of Chemistry & Center for NanoScience, Butenandtstrasse 11, Gerhard-Ertl-Gebäude, 81377 Munich, Germany

2. University of Augsburg, Institute for Physics, Universitätsstrasse 1a, 86159 Augsburg, Germany

3. Ulm University, Faculty of Natural Sciences, Institute of Biophysics Albert-Einstein Allee 11, 89081 Ulm, Germany

4. Ludwig-Maximilians-University Munich, Department of Chemistry & Center for NanoScience, Butenandtstrasse 11, Gerhard-Ertl-Gebäude, 81377 Munich, Germany.

Abstract

Aim: This study examines the absolute quantification of particle uptake into cells. Methods: We developed a novel method to analyze stacks of confocal fluorescence images of single cells interacting with nano-and micro-particles. Particle_in_Cell-3D is a freely available ImageJ macro. During the image analysis routine, single cells are reconstructed in 3D and split into two volumes – intracellular and the membrane region. Next, particles are localized and color-coded accordingly. The mean intensity of single particles, measured in calibration experiments, is used to determine the absolute number of particles. Results: Particle_in_Cell-3D was successfully applied to measure the uptake of 80-nm mesoporous silica nanoparticles into HeLa cells. Furthermore, it was used to quantify the absolute number of 100-nm polystyrene nanoparticles forming agglomerates of up to five particles; the accuracy of these results was confirmed by super-resolution, stimulated emission depletion microscopy. Conclusion: Particle_in_Cell-3D is a fast and accurate method that allows the quantification of particle uptake into cells. Original submitted 10 May 2011; Revised submitted 15 October 2012; Published online 5 February 2013

Publisher

Future Medicine Ltd

Subject

Development,General Materials Science,Biomedical Engineering,Medicine (miscellaneous),Bioengineering

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