Ekaterina Borisova, Tsanislava Genova, Latchezar Avramov
Institute of Electronics, Bulgarian Academy of Sciences, 72, Tsarigradsko chaussee Blvd.,1784 Sofia, Bulgaria
Nikolay Penkov, Ivan Terziev, Borislav Vladimirov
University Hospital "Tsaritsa Yoanna - ISUL", 8, Byalo more str., 1572 Sofia, Bulgaria
Biology Department, Saratov State University, Physiology of Human and Animals lab., 83 Astrakhanskaya str., Saratov, Russia
Synchronous fluorescence spectroscopy (SFS) is a steady-state approach that we used for evaluation of specific fluorescence characteristics of cancerous colorectal tissues. SFS allow narrowing of the fluorescence spectra received, which increase the spectral resolution and improve the analysis of the fluorescence origin in such complex onjects, such as biological tissues.
In our study we investigate the characteristic differences, with diagnostic meaning, in the synchronous fluorescence spectra (SFS) of cancerous and healthy colorectal tissues ex vivo using a spectrofluorimeter FluoroLog3 (HORIBA, JobinYvon, France) for obtaining of the SFS data in a broad spectral range (300-800 nm) using excitation in the range of 280-440 nm with a delta lambda between 0 and 200 nm with a 10 nm step between scanning excitation and emission data.
The procedure of obtaining the investigated samples ex vivo includes their excision during surgery for removal of neoplasia lesions. After the surgical removal biological samples are transported in isothermal conditions and safe-keeping solution from the hospital to the spectral laboratory, where their spectral properties were investigated. All patients received and signed written informed consent and this research is approved by Ethics committee of University Hospital “Tsaritsa Yoanna”, Sofia. Histological analysis was used as “gold standard” for evaluation of tissue samples and comparison of the spectral data received.
The main observed fluorophores which fluorescence has a diagnostic meaning are tyrosine, tryptophan, NADH, FAD, collagen, elastin, and porphyrines. Using SFS data of multiple colorectal tumours and healthy tissue originated from one patient we observed as well a formation of alterations in the fluorescence, which can be addressed to specific dysplastic changes in the tissues and could be used for tumour stage evaluation. Spectral results correlated with the histological diagnosis of the lesions investigated.
Acknowledgements: This work is supported in part by the National Science Fund of Bulgarian Ministry of Education and Science under grant #DFNI-B02/9/2014 “Development of biophotonics methods as a basis of oncology theranostics”, and in part under COST Action BM1205 “European Network for Skin Cancer Detection Using Laser Imaging”. E. Borisova and Ts. Genova personally acknowledged WFS National Scholarship Programme support and Program for support of the young scientists in Bulgarian Academy of Sciences for funding within grant \# DFNP-183- A1.
Dr. Ekaterina Georgieva Borisova
Institute of Electronics, Bulgarian Academy of Sciences, Associate Professor
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