title: Mapping the intracellular distribution of carbon nanotubes after targeted delivery to carcinoma cells using confocal Raman imaging as a label-free technique.
abstract: The uptake of carbon nanotubes (CNTs) by mammalian cells and their distribution within cells is being widely studied in recent years due to their increasing use for biomedical purposes. The two main imaging techniques used are confocal fluorescence microscopy and transmission electron microscopy (TEM). The former, however, requires labeling of the CNTs with fluorescent dyes, while the latter is a work-intensive technique that is unsuitable for in situ bio-imaging. Raman spectroscopy, on the other hand, presents a direct, straightforward and label-free alternative. Confocal Raman microscopy can be used to image the CNTs inside cells, exploiting the strong Raman signal connected to different vibrational modes of the nanotubes. In addition, cellular components, such as the endoplasmic reticulum and the nucleus, can be mapped. We first validate our method by showing that only when using the CNTs' G band for intracellular mapping accurate results can be obtained, as mapping of the radial breathing mode (RBM) only shows a small fraction of CNTs. We then take a closer look at the exact localization of the nanotubes inside cells after folate receptor-mediated endocytosis and show that, after 8-10 h incubation, the majority of CNTs are localized around the nucleus. In summary, Raman imaging has enormous potential for imaging CNTs inside cells, which is yet to be fully realized.
authors.affiliation: Institute of Biophysics, J Kepler University, Linz, Austria. constanze.lamprecht@gmx.de
authors.forename: C
authors.initials: C
authors.lastname: Lamprecht
authors.forename: N
authors.initials: N
authors.lastname: Gierlinger
authors.forename: E
authors.initials: E
authors.lastname: Heister
authors.forename: B
authors.initials: B
authors.lastname: Unterauer
authors.forename: B
authors.initials: B
authors.lastname: Plochberger
authors.forename: M
authors.initials: M
authors.lastname: Brameshuber
authors.forename: P
authors.initials: P
authors.lastname: Hinterdorfer
authors.forename: S
authors.initials: S
authors.lastname: Hild
authors.forename: A
authors.initials: A
authors.lastname: Ebner
mesh_terms: D045744:Cell Line, Tumor; D003593:Cytoplasm / Q000378:metabolism; D004705:Endocytosis; D005492:Folic Acid / Q000378:metabolism; D058972:Folic Acid Transporters / Q000378:metabolism; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism*; D018613:Microscopy, Confocal / Q000379:methods*; D037742:Nanotubes, Carbon / Q000737:chemistry; D010084:Oxidation-Reduction; D011092:Polyethylene Glycols / Q000737:chemistry; D013059:Spectrum Analysis, Raman; D001749:Urinary Bladder Neoplasms / Q000473:pathology*
chemical_list: D058972:Folic Acid Transporters; D037742:Nanotubes, Carbon; D011092:Polyethylene Glycols; D005492:Folic Acid