Technical Reference #1886

1886.

Cellular uptake and transport of gold nanoparticles incorporated in a liposomal carrier Devika B. Chithrani; Michael Dunne; James Stewart; Christine Allen; David A. Jaffray, University Health Network, Nanomedicine: Nanotechnology; Biology; and Medicine, 6(1886), (2010)
Link To Paper

Keywords:
Cell uptake; Gold nanoparticles; Intracellular transport; Liposomes

Materials & Methods:
For four-dimensional analysis of the cellular uptake of Au NP–liposomes HeLa cells were grown in glass-bottom microwell dishes (MatTek Corporation Ashland Massachusetts). Cells were incubated with Au NP–liposomes labeled with a fluorescent lipid with excitation/emission at 550/590 nm (lissamine rhodamine) for 30 minutes for their initial internalization. To clearly distinguish the individual vesicles containing Au NP–liposomes the concentration of liposomes in the media was reduced by a factor of 100 from the method described in the previous section to 1 × 107 liposomes/mL. The dish was placed on the stand of the microscope (Zeiss Two-Photon Confocal Microscope LSM 510 Meta Göttingen Germany) in such a way that the coverslip area was in contact with the × 63 waterimmersed microscope objective. Sequences of Z-stacks were taken for a group of selected cells in 5-minute time intervals for real-time analysis of transport within the cell cytoplasm. Cell permeable Lyso Tracker (Lyso Tracker Green DND-26; Molecular Probes-L7526 Eugene Oregon) was employed for live cell staining of lysosomes. To study the distribution of the Au NP–liposomes in organelles the cells were incubated with 60 μL Lyso Tracker (50 nM) (excitation/emission at 504/511 nm) along with the liposomes labeled with rhodamine fluorescent dye (excitation/emission at 550/590 nm) for 30 minutes.

Microscopic Technique
Confocal Microscopy

Cell Type(s)
HeLa cells