Analysis of oxidized multi-walled carbon nanotubes in single K562 cells by capillary electrophoresis with laser-induced fluorescence.
Short oxidized multi-walled carbon nanotubes (CNT) were derivatized with fluorescein isothiocyanate (FITC). Capillary electrophoresis coupled with laser-induced fluorescence (CE-LIF) was then used to separate and detect the fluorescently labeled carbon-nanotube probes (CNTP) in multidrug-resistant cells (K562A) and the parent cells (K562S). Greater expression of P-glycoprotein in K562A cells than in K562S cells was confirmed by use of anti-P-glycoprotein antibody and flow-cytometric analysis. Analyses of CNTP in both cell lines using both CE-LIF and flow cytometry showed that CNTP could traverse the cellular membrane without being pumped out by P-glycoprotein. The CNTP distributed in both cell lines was analyzed at the single cell level and the results were compared with those from analysis of ten cells and of the lysate from bulk cells. The results revealed the CE-LIF method could be used for quantitative analysis of CNT in single cells in studies of drug delivery and multidrug resistance.
Xiao
Hua
H
National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, People's Republic of China.
Yang
Lisong
L
Zou
Hanfa
H
Yang
Ling
L
Le
X Chris
XC
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000032:analysis / Q000378:metabolism; D019075:Electrophoresis, Capillary / Q000379:methods*; D004705:Endocytosis; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D006801:Humans; D020014:K562 Cells; D007834:Lasers; D046529:Microscopy, Electron, Transmission; D037742:Nanotubes, Carbon / Q000032:analysis* / Q000737:chemistry / Q000648:ultrastructure; D010084:Oxidation-Reduction; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods*; D013696:Temperature
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D037742:Nanotubes, Carbon; D012967:Sodium Dodecyl Sulfate; D016650:Fluorescein-5-isothiocyanate