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

作者：Torrano Adriano A; Blechinger Julia; Osseforth Christian; Argyo Christian; Reller Armin; Bein Thomas; Michaelis Jens; Braeuchle Christoph

来源：Nanomedicine, 2013, 8(11): 1815-1828.

DOI：10.2217/nnm.12.178

• cellular uptake
• fluorescence microscopy
• ImageJ
• nanomedicine
• nanoparticle
• nanotoxicology
• STED
• super-resolution microscopy

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

• 出版日期2013-11