Imaging receptor-mediated endocytosis with a polymeric nanoparticle-based coherent anti-stokes Raman scattering probe.
Coherent anti-Stokes Raman scattering (CARS) microscopy was used to visualize receptor-mediated endocytosis and intracellular trafficking with the aid of a CARS probe. The probe was made of 200-nm polystyrene particles encapsulated in folate-targeted liposomes. By tuning (omega(p) - omega(s)) to 3045 cm(-1), which corresponds to the aromatic C-H stretching vibration, the polystyrene nanoparticles with a high density of aromatic C-H bonds were detected with a high signal-to-noise ratio, while the epi-detected CARS signal from cellular organelles was cancelled by the destructive interference between the resonant contribution from the aliphatic C-H vibration and the nonresonant contribution. Without any photobleaching, the CARS probe allowed single-particle tracking analysis of intracellular endosome transport. No photodamage to cells was observed under the current experimental conditions. These results show the advantages and potential of using a CARS probe to study cellular processes.
Department of Chemistry, Purdue University, West Lafayette, Indiana 47906, USA.
Ling
L
Tong
Yanhui
Y
Lu
Robert J
RJ
Lee
Ji-Xin
JX
Cheng
D003078:Colchicine / Q000494:pharmacology; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology*; D005452:Fluoresceins / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D007624:KB Cells; D007834:Lasers; D008956:Models, Chemical; D053758:Nanoparticles; D011137:Polystyrenes / Q000737:chemistry; D011956:Receptors, Cell Surface / Q000502:physiology*; D012542:Scattering, Radiation; D013059:Spectrum Analysis, Raman
D005452:Fluoresceins; D011137:Polystyrenes; D011956:Receptors, Cell Surface; D003078:Colchicine; C007740:fluorexon