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Live‐Cell Imaging by Confocal Raman and Fluorescence Microscopy Recognizes the Crystal Structure of Calcium Carbonate Particles in HeLa Cells
Author(s) -
Abalymov Anatolii A.,
Verkhovskii Roman A.,
Novoselova Marina V.,
Parakhonskiy Bogdan V.,
Gorin Dmitry A.,
Yashchenok Alexey M.,
Sukhorukov Gleb B.
Publication year - 2018
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201800071
Subject(s) - vaterite , calcite , biophysics , calcium carbonate , raman spectroscopy , confocal , confocal microscopy , fluorescence , biomineralization , fluorescence microscope , cytoplasm , chemistry , materials science , chemical engineering , mineralogy , biochemistry , biology , microbiology and biotechnology , optics , physics , organic chemistry , engineering , aragonite
Porous calcium carbonate (CaCO 3 ) vaterite particles are very attractive templates for the encapsulation of pharmaceuticals and for the construction of hollow polyelectrolyte capsules, sensors, and enzyme‐catalyzed reactors. Although CaCO 3 is biocompatible and biodegradable, little is known about the intercellular behavior and properties of vaterite particles in the cytoplasm of cells. In this work, the authors combine confocal Raman and fluorescent microscopy for the imaging of porous CaCO 3 vaterite particles in HeLa cells to study the uptake and status of the particles inside the cells in real time. Analysis of the fluorescence images shows that the particles penetrated the plasma membrane 3 h after being added to the cell culture and that the internalization of the particles continued up to 48 h. The crystal structure of individual vaterite particles in the cytoplasm of HeLa cells did not obviously change for 144 h. For clusters of particles, however, the authors identify Raman spectroscopic signatures of the stable calcite phase after 72 h of incubation, confirming an ion‐exchange mechanism of vaterite transformation to calcite. The results indicate that these imaging approach to examining inorganic particles in living cells may have theranostic applications.