Phototherapy of Brain Tumours Using a Fibre Optic Neurosystem

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.

State transitions and photosystems spatially resolved in individual cells of the cyanobacterium Synechococcus elongatus

State transitions are a low-light acclimation response through which the excitation of Photosystem I (PSI) and Photosystem II (PSII) is balanced; however, our understanding of this process in cyanobacteria remains poor. Here, picosecond fluorescence kinetics was recorded for the cyanobacterium Synechococcus elongatus using fluorescence lifetime imaging microscopy (FLIM), both upon chlorophyll a and phycobilisome (PBS) excitation. Fluorescence kinetics of single cells obtained using FLIM were compared with those of ensembles of cells obtained with time-resolved fluorescence spectroscopy. The global distribution of PSI and PSII and PBSs was mapped making use of their fluorescence kinetics. Both radial and lateral heterogeneity were found in the distribution of the photosystems. State transitions were studied at the level of single cells. FLIM results show that PSII quenching occurs in all cells, irrespective of their state (I or II). In S. elongatus cells, this quenching is enhanced in State II. Furthermore, the decrease of PSII fluorescence in State II was homogeneous throughout the cells, despite the inhomogeneous PSI/PSII ratio. Finally, some disconnected PBSs were resolved in most State II cells. Taken together our data show that PSI is enriched in the inner thylakoid, while state transitions occur homogeneously throughout the cell.

Определение констант скоростей и механизма фотопревращений металлопорфиринов методом решения обратной фотокинетической задачи

The fluorescence kinetics of some substituted Zn- and Mg-porphyrins in solid polymer films upon excitation by two-step-wise rectangular laser pulses and 293 K were investigated by the inverse kinetic problem methods. The experimental non-monotonic kinetic curves were approximated by the simulated ones within the framework of a six-level energy scheme describing the reversible interconversions between the two complexes. The best fit between the experimental and simulated curves was obtained by iterative optimization using the Nelder-Mead algorithm. Based on the statistically estimated values ​​of the rate constants and parameters of the considered models, an interpretation of the observed kinetics was given and the conclusion drawn that the interconversions are caused by the process of reversible extra-liganding of the central metal ion, which occurs in the excited triplet T₁ state.

Chlorophyll Fluorescence Kinetics May Be Useful to Identify Early Drought and Irrigation Effects on Photosynthetic Apparatus in Field-Grown Wheat

To assess the reliability and sensitivity of non-invasive optical methods to detect the early effects of water deficit in the field, we analyzed the time-series of non-invasive measurements obtained in a dry season in a representative collection of wheat genotypes grown in small-plot field trials, in non-irrigated and irrigated variants. Despite a progressive water deficit and significant yield loss, the measurements indicated very minor changes in chlorophyll content or canopy cover. This corresponded well to the insignificant differences in spectral reflectance normalized difference vegetation index (NDVI) values. On the other hand, we identified the significant and rapid response of fast fluorescence kinetics data following the onset of irrigation. Analysis of parameters showed the main effects of drought were associated with changes in the amplitude of the I–P phase of the OJIP transient, indicating changes at the level of photosystem I and beyond. Statistical analyses identified the integrative parameter performance index PItot as the most sensitive parameter, which well-reflects the differences in responses of the genotypes to water deficit. Our results suggest that focusing on photosynthetic functions detected by the rapid chlorophyll fluorescence records can provide more accurate information on the drought stress level, compared to the structural data obtained by absorbance or reflectance measurements.