Quantitative Mapping of the Spatial Distribution of Nanoparticles in Endo-Lysosomes by Local pH.
Jing WangSarah R MacEwanAshutosh ChilkotiPublished in: Nano letters (2017)
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.
Keyphrases
- drug delivery
- fluorescence imaging
- cancer therapy
- drug release
- quantum dots
- induced apoptosis
- high resolution
- photodynamic therapy
- living cells
- cell cycle arrest
- reactive oxygen species
- stem cells
- magnetic resonance imaging
- cell therapy
- single cell
- hydrogen peroxide
- magnetic resonance
- walled carbon nanotubes
- high density
- bone marrow
- emergency department
- electronic health record
- molecularly imprinted
- simultaneous determination
- liquid chromatography
- loop mediated isothermal amplification