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Dynamic colocalization microscopy to characterize intracellular trafficking of nanomedicines

(2011) ACS NANO. 5(10). p.7874-7884
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Abstract
To gain a better understanding of intracellular processing of nanomedicines, we employed quantitative live-cell fluorescence colocalization microscopy to study endosomal trafficking of polyplexes in retinal pigment epithelium cells. A new, dynamic colocalization algorithm was developed, based on particle tracking and trajectory correlation, allowing for spatiotemporal characterization of internalized polyplexes in comparison with endosomal compartments labeled with EGFP constructs. This revealed early trafficking of the polyplexes specifically to Rab5- and flotillin-2-positive vesicles and subsequent delivery to Rab7 and LAMP1-labeled late endolysosomes where the major fraction of the polyplexes remains entrapped for days, suggesting the functional loss of these nanomedicines. Colocalization of polyplexes with the autophagy marker LC3 suggests for the first time that the process of xenophagy could play an important role in the persistent endosomal entrapment of nanomedicines.
Keywords
particle tracking, AUTOPHAGY, PLASMID DNA, DRUG, intracellular trafficking, nonviral ocular gene therapy, nanomedicines, colocalization, autophagy, GENE DELIVERY, FLUORESCENCE MICROSCOPY, SINGLE MOLECULES, LIVING CELLS, PARTICLE TRACKING, LYSOSOMES, TRANSPORT

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MLA
Vercauteren, Dries, Hendrik Deschout, Katrien Remaut, et al. “Dynamic Colocalization Microscopy to Characterize Intracellular Trafficking of Nanomedicines.” ACS NANO 5.10 (2011): 7874–7884. Print.
APA
Vercauteren, Dries, Deschout, H., Remaut, K., Engbersen, J. F., Jones, A. T., Demeester, J., De Smedt, S., et al. (2011). Dynamic colocalization microscopy to characterize intracellular trafficking of nanomedicines. ACS NANO, 5(10), 7874–7884.
Chicago author-date
Vercauteren, Dries, Hendrik Deschout, Katrien Remaut, Johan FJ Engbersen, Arwyn T Jones, Jo Demeester, Stefaan De Smedt, and Kevin Braeckmans. 2011. “Dynamic Colocalization Microscopy to Characterize Intracellular Trafficking of Nanomedicines.” Acs Nano 5 (10): 7874–7884.
Chicago author-date (all authors)
Vercauteren, Dries, Hendrik Deschout, Katrien Remaut, Johan FJ Engbersen, Arwyn T Jones, Jo Demeester, Stefaan De Smedt, and Kevin Braeckmans. 2011. “Dynamic Colocalization Microscopy to Characterize Intracellular Trafficking of Nanomedicines.” Acs Nano 5 (10): 7874–7884.
Vancouver
1.
Vercauteren D, Deschout H, Remaut K, Engbersen JF, Jones AT, Demeester J, et al. Dynamic colocalization microscopy to characterize intracellular trafficking of nanomedicines. ACS NANO. 2011;5(10):7874–84.
IEEE
[1]
D. Vercauteren et al., “Dynamic colocalization microscopy to characterize intracellular trafficking of nanomedicines,” ACS NANO, vol. 5, no. 10, pp. 7874–7884, 2011.
@article{2134082,
  abstract     = {{To gain a better understanding of intracellular processing of nanomedicines, we employed quantitative live-cell fluorescence colocalization microscopy to study endosomal trafficking of polyplexes in retinal pigment epithelium cells. A new, dynamic colocalization algorithm was developed, based on particle tracking and trajectory correlation, allowing for spatiotemporal characterization of internalized polyplexes in comparison with endosomal compartments labeled with EGFP constructs. This revealed early trafficking of the polyplexes specifically to Rab5- and flotillin-2-positive vesicles and subsequent delivery to Rab7 and LAMP1-labeled late endolysosomes where the major fraction of the polyplexes remains entrapped for days, suggesting the functional loss of these nanomedicines. Colocalization of polyplexes with the autophagy marker LC3 suggests for the first time that the process of xenophagy could play an important role in the persistent endosomal entrapment of nanomedicines.}},
  author       = {{Vercauteren, Dries and Deschout, Hendrik and Remaut, Katrien and Engbersen, Johan FJ and Jones, Arwyn T and Demeester, Jo and De Smedt, Stefaan and Braeckmans, Kevin}},
  issn         = {{1936-0851}},
  journal      = {{ACS NANO}},
  keywords     = {{particle tracking,AUTOPHAGY,PLASMID DNA,DRUG,intracellular trafficking,nonviral ocular gene therapy,nanomedicines,colocalization,autophagy,GENE DELIVERY,FLUORESCENCE MICROSCOPY,SINGLE MOLECULES,LIVING CELLS,PARTICLE TRACKING,LYSOSOMES,TRANSPORT}},
  language     = {{eng}},
  number       = {{10}},
  pages        = {{7874--7884}},
  title        = {{Dynamic colocalization microscopy to characterize intracellular trafficking of nanomedicines}},
  url          = {{http://dx.doi.org/10.1021/nn2020858}},
  volume       = {{5}},
  year         = {{2011}},
}

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