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Fluorescence microspectroscopy as a tool to study mechanism of nanoparticles delivery into living cancer cells
Authors:Arsov Zoran  Urbančič Iztok  Garvas Maja  Biglino Daniele  Ljubetič Ajasja  Koklič Tilen  Strancar Janez
Affiliation:1Laboratory of Biophysics, Department of Solid State Physics, Jožef Stefan Institute, Jamova 39, 1000 Ljubljana, Slovenia;2Center of Excellence NAMASTE, Jamova 39, 1000 Ljubljana, Slovenia
Abstract:Lack of better understanding of nanoparticles targeted delivery into cancer cells calls for advanced optical microscopy methodologies. Here we present a development of fluorescence microspectroscopy (spectral imaging) based on a white light spinning disk confocal microscope with emission wavelength selection by a liquid crystal tunable filter. Spectral contrasting of images was used to localize polymer nanoparticles and cell membranes labeled with fluorophores that have substantially overlapping spectra. In addition, fluorescence microspectroscopy enabled spatially-resolved detection of small but significant effects of local molecular environment on the properties of environment-sensitive fluorescent probe. The observed spectral shift suggests that the delivery of suitably composed cancerostatic alkylphospholipid nanoparticles into living cancer cells might rely on the fusion with plasma cell membrane.
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