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Report

Teaser, summary, work performed and final results

Periodic Reporting for period 1 - FORECAST (Fluorescence lifetime optical biopsy system)

Teaser

Summary

The â€œFluorescence lifetime optical biopsy systemâ€ (FORECAST) Fellowship has successfully reached its ultimate goal of training a talented researcher, Dr. Evgeny Zherebtsov, through a research project in the area of developing laser-based photonic devices for biomedical research and clinical diagnostics. The research in the area of optical biopsy systems, where fluorescence spectroscopy is one of the key approaches, is rapidly expanding due to its safety, relative cheapness and efficiency. The ability to recognize the suspect tissue in a second without interruption of the procedure dramatically increases the effectiveness of the endoscopic surgical removal of the cancerous tumours. The project focused on the development of novel tools for the in vivo real-time assessment of the respiratory chain activity (NADH/FAD ratio) and mitochondrial dysfunction in tissues. This is achieved by means of both laser fluorescence spectra and lifetime fluorescence data analysis supplemented with additional optical measurements (laser Doppler flowmetry, tissue reflectance spectroscopy) in order to correct the impact of affecting factors (changes in blood perfusion, blood volume, blood oxygenation).
The FORECAST project sets the following list of key objectives (RO): broadening of the Fellowâ€™s expertise in frequency domain lifetime fluorescence spectroscopy with a focus on medical applications; development of novel theoretical models for the registering fluorescence parameters in living epithelium tissue; design an advanced technique for registration of the fluorescence lifetime fluorescence intensity in a wide spectral range simultaneously with LDF and tissue blood oximetry measurements; expansion of experience in the field of the assessment of mitochondrial dysfunction according to the fluorescence parameters in tissues; expansion of experience in the field of medical monitoring for non-invasive fluorescence diagnosis of epithelium cancers.

Work performed

WP1: The overall training activities and knowledge transfer. Courses attended: COSHH assessments (6th June 2017); Human tissue act training (23rd January 2017); GCP for Laboratory staff working with anonymized samples (23rd January 2017); Risk assessments (30th May 2017); Research Funding Seminar Series: Fellowships (12th April 2017); Open Access & PURE (9th May 2017); Grant Writing Workshops (16th Jan, 13th Feb 2018), EPSRC eFutures Early Career Researcherâ€™s Workshop (13th/14th September, Murray Edwards College, Cambridge, 2018).
WP2: Development of novel theoretical models for the fluorescence lifetime and intensity in the living epithelium tissue. Work performed: modelling of diagnostic volume for the fluorescence measurements for the different excitation wavelength for the parameters of the used optical fiber probe; modelling for the diagnostic volume for the LDF measurements; theoretical studies on impact of blood volume on the diffuse reflectance spectra of human skin; analysis of the component demixing using phasor analysis of the lifetime fluorescence; development of novel theoretical model for the artificial neural network aided blood volume fraction and the oxygen saturation reconstruction. Dissemination: JBO 22 (8), 085003; Photonics West 2017; Proc. Spie 104120H; Proc. Spie 104120G; Proc. SPIE 1071619. Conferences: European Conferences on Biomedical Optics 2017; Saratov Fall Meeting 2017.
WP3: The design an advanced technique for registration of the fluorescence lifetime and intensity of the emission in the wide spectral range. Work performed: setup development for measurements of the fluorescence intensity and lifetime in frequency domain using high speed lock-in amplifier; development of the diagnostic fibre optical probe; software for the blood influence compensation. Development of the VCSEL-based blood perfusion sensor. Tests with Ce3+-doped silica fiber at different pump wavelengths. Dissemination: Laser Physics Letters 14 (6), 065603; Laser Physics Letters 15 (10), 105601; Proc. 2018 ICLO, 133-133; Proc. SPIE 104931L; Proc. of SPIE Vol. 1006303-1. Conferences: Laser Optics 2018, Photonics West 2017, Photonics West 2018, 10th ESGCO 2018 (Invited talk).
WP4: Expansion of experience in the field of medical monitoring for non-invasive fluorescence diagnosis of epithelium tissues. Research protocol development, obtaining ethical approval. Studies on the influence of the blood microflow on the registered parameters of fluorescence. Dissemination: Microvascular Research, 120, pp. 13-20, 2018; Proc. Spie 104120W; 2018 International Conference Laser Optics (ICLO), 564-564; Proc. Spie 106854C; Proc. SPIE 1068532; Proc. SPIE 106854O. Conferences: Laser Optics 2018, European Conferences on Biomedical Optics 2017, ECBO 2018.
WP5: Expansion of experience in the field of the assessment of mitochondrial dysfunction according to the fluorescence parameters in the epithelium in vitro. Work performed: testing of the developed diagnostic technology on cultures of living tissue with inhibition of components of the electron transport in the respiratory chain; verification of NADH content measurements in slices of living brain tissue. Dissemination: Proc. SPIE 106854E; Proc. SPIE 106854R. Conference: Photonics Europe 2018, Rank Prize Symposium, 2018.
WP6: Expansion of experience in the field of medical monitoring of mitochondrial dysfunction by non-invasive fluorescence diagnosis during drug discovery of muscular dystrophy treatment. Research protocol development, experiment preparations and arrangements. Due to the cutting down of the Fellowship duration to launch the next research project, the work package 6 has not been developed completely.

Final results

1) Demonstration that the intensity of skin fluorescence and level of tissue blood perfusion can act as markers for various degrees of complications from the beginning of diabetes to the formation of trophic ulcers. 2) New diagnostic criterion allowing for the relation of current tissue vitality level to one of the three groups (healthy, moderate complications, severe complications). The findings can make a significant contribution to the fight against diabetes, which is very relevant given the prevalence of the disease. 3) Justified ability to control the passage of fluorescent dye-filled liposomes through the epithelial tissues after oral administration by proposed system. A potential application is monitoring orally administered liposomal particles. The results can be used to help create new optical tools for use in the development of new drugs and in high-throughput screening used during their testing. 4) Evaluated dynamics of the dissemination of dye-filled nanocapsules in the adjacent tissues via the circulatory system which was observed and assessed quantitatively. The approach can be used for the transdermal assessment of rhodamine-loaded capsules. 5) Observed features in skin and epithelial blood microflow, which can lead to new diagnostical criteria for diagnostics of diabetes mellitus complications. 6) New approach for hyperspectral diagnostics aided by artificial neural networks for functional skin characterization. Potential applications of the proposed technique are of high social impact and include monitoring and diagnostics of diabetic ulcer formation, rheumatic complications, diagnoses of melanoma and other malignancies, quantitative skin screening, wound healing control, assistance and support of laser surgical and treatment procedures and others.