Magnetic CaCO3-protein-cyanine dyes multicomposite for co-visualization in vitro and in vivo
Alexandra E. Kalinova1, Ludmila I. Kuznetsova1, Roman A. Anisimov1, Alla B. Bucharskaya2, Maria V. Lomova1; 1Science Medical Centre, Saratov State University; 2Head of the Center for Collective Use of Economic Opportunities, Deputy Director of the Research Institute for Scientific Work, Saratov State Medical University named after V. I. Razumovsky
Abstract
Luminescent bioimaging is based on labeling the object of study (for example, a drug molecule or a cell organelle) with a luminescent label, the signal of which can be detected when excitation energy is applied. In this regard, the goal of my work was to develop a technology for obtaining a multicomposite based on CaCO3 and cyanine dyes for co-visualization in vitro and in vivo.
Pure dyes Sulfo-Cyanine5.5, 7 and 7.5 NHS esters were dissolved in alcohol and then mixed with a buffer solution. Next, 6 different ratios of dyes were used to obtain 6 complexes, which were subsequently conjugated with bovine serum albumin (BSA. The absorption spectra and fluorescence were evaluated using a spectrophotometer CLARIOstar (Offenburg, Germany). Fluorescence quenching was shown to occur by a dynamic mechanism due to random collisions of protein molecules with fluorophore molecules.
Next, the fluorescence of samples was assessed using a bioluminograph Fluor I In Vivo (NeoScience, Daejeon, South Korea). To obtain a multicomposite, submicron vaterite was synthesized, loaded with magnetite nanoparticles, and conjugates with protein were adsorbed onto the resulting capsules. Red channel with a sNIR filter, in the NIR channel with a NIR filter and in the green channel with a red filter were used.
The final stage of the study was flow cytometry of pure dyes Cy5.5 and Cy7 and complexes Cy5.5(2):Cy7(1), Cy5.5(1):Cy7(2).
The work was carried out within the framework of the RSF project 23-13-00373 «Mechanism of antitumor action of an alternating non-heating magnetic field in vitro and in vivo».
Speaker
Alexandra E. Kalinova
Science Medical Centre, Saratov State University
Russia
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