A fast analysis method to quantify nanoparticle uptake on a single cell level. (November 2013)

• Record Type: Journal Article
• Authors: Torrano, Adriano A; Blechinger, Julia; Osseforth, Christian; Argyo, Christian; Reller, Armin; Bein, Thomas; Michaelis, Jens; Bräuchle, Christoph
• Is Part Of: Nanomedicine. Volume 8:Number 11(2013)
• Page Start: 1815
• Page End: 1828
• Publication Date: 2013-11
• Subjects: cellular uptake -- fluorescence microscopy -- ImageJ -- nanomedicine -- nanoparticle -- nanotoxicology -- STED -- super-resolution microscopy; Nanotechnology -- Periodicals; Medical technology -- Periodicals; Nanotechnology -- Therapeutic use -- Periodicals; 610.28
• DOI: 10.2217/nnm.12.178
• Languages: English
• ISSNs: 1743-5889

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