title: Quantitative Mapping of the Spatial Distribution of Nanoparticles in Endo-Lysosomes by Local pH.
abstract: Understanding the intracellular distribution and trafficking of nanoparticle drug carriers is necessary to elucidate their mechanisms of drug delivery and is helpful in the rational design of novel nanoparticle drug delivery systems. The traditional immunofluorescence method to study intracellular distribution of nanoparticles using organelle-specific antibodies is laborious and subject to artifacts. As an alternative, we developed a new method that exploits ratiometric fluorescence imaging of a pH-sensitive Lysosensor dye to visualize and quantify the spatial distribution of nanoparticles in the endosomes and lysosomes of live cells. Using this method, we compared the endolysosomal distribution of cell-penetrating peptide (CPP)-functionalized micelles to unfunctionalized micelles and found that CPP-functionalized micelles exhibited faster endosome-to-lysosome trafficking than unfunctionalized micelles. Ratiometric fluorescence imaging of pH-sensitive Lysosensor dye allows rapid quantitative mapping of nanoparticle distribution in endolysosomes in live cells while minimizing artifacts caused by extensive sample manipulation typical of alternative approaches. This new method can thus serve as an alternative to traditional immunofluorescence approaches to study the intracellular distribution and trafficking of nanoparticles within endosomes and lysosomes.
authors.affiliation: Department of Biomedical Engineering, Duke University , Durham, North Carolina 27708, United States.
authors.forename: Jing
authors.identifier: 0000-0001-8264-4405
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Biomedical Engineering, Duke University , Durham, North Carolina 27708, United States.
authors.forename: Sarah R
authors.initials: SR
authors.lastname: MacEwan
authors.affiliation: Department of Biomedical Engineering, Duke University , Durham, North Carolina 27708, United States.
authors.forename: Ashutosh
authors.initials: A
authors.lastname: Chilkoti
mesh_terms: D002460:Cell Line; D057846:Cell-Penetrating Peptides / Q000737:chemistry; D004337:Drug Carriers / Q000737:chemistry; D011992:Endosomes / Q000378:metabolism; D005456:Fluorescent Dyes; D006801:Humans; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D008247:Lysosomes / Q000378:metabolism*; D008823:Micelles; D053758:Nanoparticles / Q000737:chemistry*; D061848:Optical Imaging; D013499:Surface Properties
chemical_list: D057846:Cell-Penetrating Peptides; D004337:Drug Carriers; D005456:Fluorescent Dyes; D008823:Micelles
keywords: Fluorescence imaging; cell-penetrating peptide; lysosome; nanoparticle