28 February 2014 The study of radiation-induced damage and remodeling of extracellular matrix of rectum and bladder by second-harmonic generation microscopy
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Adverse events in normal tissues after irradiation of malignant tumors are of great importance in modern radiation oncology. Second harmonic generation (SHG) microscopy allows observe the structure of collagen fibers and bundles without additional staining. The study objective was evaluation the dose-time dependences of the structural changes occurring in collagen of rat rectum and bladder after gamma-irradiation. Animals were irradiated by a local field at single doses of 10 Gy and 40 Gy. The study of collagen state was carried out in a week and a month after radiation exposure. Paraffin-embedded material was sectioned on the slices 10 mkm thick and SHG-imaging was performed by LSM 510 Meta (Carl Zeiss, Germany). Excitation was implemented with a pulsed (100-fs) titanium-sapphire laser at a wavelength of 800 nm and a pulse repetition frequency of 80 MHz, registration was performed at two wavelengths: 362-415 nm according to collagen fluorescence and 512-576 nm according to myoglobin fluorescence. In a week after irradiation, sings of epithelial damage and edema of submucosal layer, more significant after the dose of 40 Gy were observed on LSM-images. The SHG signal decreased at this time reflecting the processes of collagen degradation independently either in bladder or in rectum. In a month after radiation the increase of size and number of collagen-bearing structures was observed, more essential after irradiation in a dose of 40 Gy. LSM microscopy with SHG allows evaluate changes of normal tissues after ionizing radiation and get information in addition to standard and special histological staining.
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Marina V. Kochueva, Marina V. Kochueva, Ekaterina A. Sergeeva, Ekaterina A. Sergeeva, Natalya Yu. Ignatjeva, Natalya Yu. Ignatjeva, Olga L. Zakharkina, Olga L. Zakharkina, Sergej S. Kuznetzov, Sergej S. Kuznetzov, Elena B. Kiseleva, Elena B. Kiseleva, Ksenia V. Babak, Ksenia V. Babak, Vladislav A. Kamensky, Vladislav A. Kamensky, Anna V. Maslennikova, Anna V. Maslennikova, "The study of radiation-induced damage and remodeling of extracellular matrix of rectum and bladder by second-harmonic generation microscopy", Proc. SPIE 8948, Multiphoton Microscopy in the Biomedical Sciences XIV, 894809 (28 February 2014); doi: 10.1117/12.2039035; https://doi.org/10.1117/12.2039035

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