Title:Biological Efficacy of Ionizing Radiation Sources on 3D Organotypic Tissue Slices Assessed by Fluorescence Microscopy
Volume: 18
Issue: 1
Author(s): Victoria Shestakova, Anna Smirnova, Anna Yakimova, Sergey Koryakin, Denis Baranovskii, Vyacheslav Saburov, Elena Yatsenko, Alexander Moiseev, Sergey Ivanov, Ekaterina Smirnova, Svetlana Belkina, Grigory Demyashkin, Lyudmila Komarova, Peter Shegay, Andrey Kaprin and Ilya Klabukov*
Affiliation:
- Department of Regenerative Medicine, National Medical Research Radiological Center of the Ministry of Health of
the Russian Federation, Obninsk, Russia
- Obninsk Institute for Nuclear Power Engineering of the National Research
Nuclear University MEPhI, Obninsk, Russia
- Patrice Lumumba Peoples’ Friendship University of Russia (RUDN
University), Moscow, Russia
Keywords:
cultured viable tissue slices, ionizing radiation, fluorescent microscopy, fluorescent staining, radiobiology.
Abstract:
Objective: Traditional cell-based radiobiological methods are inadequate for assessing
the toxicity of ionizing radiation exposure in relation to the microstructure of the extracellular
matrix. Organotypic tissue slices preserve the spatial organization observed in vivo,
making the tissue easily accessible for visualization and staining. This study aims to explore the
use of fluorescence microscopy of physiologically relevant 3D tissue cultures to assess the effects
of ionizing radiation.
Methods: Organotypic tissue slices were obtained by vibratome, and their mechanical properties
were studied. Slices were exposed by two ionizing radiation sources; electron beams (80 Gy
and 4 Gy), and soft gamma irradiation (80 Gy and 4 Gy). Two tissue culture protocols were
used: the standard (37°C), and hypothermic (30°C) conditions. A qualitative analysis of cell
viability in organotypic tissue slices was performed using fluorescent dyes and standard laser
confocal microscopy.
Results: Biological dosimetry is represented by differentially stained 200-μm thick organotypic
tissue sections related to living and dead cells and cell metabolic activity.
Conclusion: Our results underscore the ability of fluorescence laser scanning confocal microscopy
to rapidly assess the radiobiological effects of ionizing radiation in vitro on 3D organotypic
tissue slices.
