Journal
NANOMEDICINE
Volume 8, Issue 11, Pages 1815-1828Publisher
FUTURE MEDICINE LTD
DOI: 10.2217/nnm.12.178
Keywords
cellular uptake; fluorescence microscopy; ImageJ; nanomedicine; nanoparticle; nanotoxicology; STED; super-resolution microscopy
Funding
- Deutsche Forschungsgemeinschaft (DFG) [SPP 1313]
- Nanosystems Initiative Munich (NIM)
- Center for Integrated Protein Science Munich (CIPSM)
- BioImaging Network (BIN) Munich
- International Doctorate Program NanoBioTechnology (IDK-NBT)
- Romer Foundation
- European Research Council
- [SFB 749]
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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
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