Abstract
In this work, a new approach was tested to assess the cellular composition of tissues by time-resolved methods of fluorescence analysis of exogenous and endogenous fluorophores. First of all, the differences in fluorescence kinetics of endogenous fluorophores (coenzymes NADH and FAD) in tumour and immunocompetent cells were determined. After that, differences in fluorescence kinetics of photosensitizer 5 ALA-induced protoporphyrin IX were established due to its different metabolism in cells of different phenotypes. Kinetics of photoluminescence of NADH and FAD coenzymes as well as photosensitizer were studied by means of two different methods: time-resolved spectroscopy based on a streak-camera and fibre optic neuroscopy, which served to perform process monitoring and regular fluorescence diagnosis of the probed region. Time-resolved fluorescence microscopy (FLIM) was used as a control technique. Time-resolved spectroscopic fluorescence lifetime analysis was performed on sexually mature female rats induced with glioma C6 brain tumour under in vivo conditions; thus, under conditions where the immune system actively intervenes in the process of oncogenesis. In this regard, the aim of the study was to recognize the cellular composition of the brain tumour tissue, namely the ratio of cancer and immunocompetent cells and their mutual localization. Understanding the role of the immune system thus provides new ways and approaches for further diagnosis and therapy, making tumour-associated immune cells a prime target for modern therapies.
Highlights
Especially time-resolved optical spectroscopy, for the analysis of brain condition and function have significant advantages over other techniques used in neurosurgery and neuro-oncology [1,2,3,4]
Time-resolved spectroscopy and microscopy make it possible to assess, by recording the fluorescence lifetime of externally injected photosensitizers, the condition of tissues that differ in phenotype due to their different metabolism and the nature of interaction with photosensitizer molecules [8,9,10]
Be established how the quantitative of tumour-associated it will itbewill established how the quantitative ratio ofratio tumour-associated macromacrophages/microglia influences the pattern of tumour progression
Summary
Especially time-resolved optical spectroscopy, for the analysis of brain condition and function have significant advantages over other techniques used in neurosurgery and neuro-oncology [1,2,3,4]. Important is the wide range of physiological and morphological parameters available for optical spectroscopy analysis. These methods allow for a clear correlation between the recorded signal—absorption, fluorescence or different types of scattering due to both substances originally inherent to neural tissues and cells and contrasting markers introduced from outside—and small and rapid functional changes in the examined tissues, and show a strong correlation with the profound metabolic and structural rearrangements that take place in the development of brain pathologies [5,6,7]. A neurosystem with an intracranial fibre structure has been previously developed that enables one to deliver laser radiation to a considerable depth inside the brain tissue after its intracranial implantation [11]
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