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Intracellular dynamics of topoisomerase I inhibitor,CPT-11, by slit-scanning confocal Raman microscopy
Authors:Yoshinori Harada  Ping Dai  Yoshihisa Yamaoka  Mitsugu Ogawa  Hideo Tanaka  Kazuto Nosaka  Kenichi Akaji  Tetsuro Takamatsu
Institution:(1) Department of Pathology and Cell Regulation, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, 465 Kajii-cho, Hirokoji Kawaramachi, Kamigyo-ku, Kyoto 602-8566, Japan;(2) Department of Chemistry, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Daishogun, Kita-ku, Kyoto 603-8334, Japan
Abstract:Most molecular imaging technologies require exogenous probes and may have some influence on the intracellular dynamics of target molecules. In contrast, Raman scattering light measurement can identify biomolecules in their innate state without application of staining methods. Our aim was to analyze intracellular dynamics of topoisomerase I inhibitor, CPT-11, by using slit-scanning confocal Raman microscopy, which can take Raman images with high temporal and spatial resolution. We could acquire images of the intracellular distribution of CPT-11 and its metabolite SN-38 within several minutes without use of any exogenous tags. Change of subcellular drug localization after treatment could be assessed by Raman imaging. We also showed intracellular conversion from CPT-11 to SN-38 using Raman spectra. The study shows the feasibility of using slit-scanning confocal Raman microscopy for the non-labeling evaluation of the intracellular dynamics of CPT-11 with high temporal and spatial resolution. We conclude that Raman spectromicroscopic imaging is useful for pharmacokinetic studies of anticancer drugs in living cells. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.
Keywords:Slit-scanning Raman microscopy  Molecular imaging  Non-labeling method  Anticancer-drug
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