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Biomedical subjects

A Rück

Publications and source records attributed to A Rück.

45 records · Page 3Linked to original sources
PubMed1992

Intracellular localization of meso-tetraphenylporphine tetrasulphonate probed by time-resolved and microscopic fluorescence spectroscopy.

The effects of solvent pH on spectral properties and fluorescence decay kinetics were investigated in order to characterize the microenvironment of meso-tetraphenylporphine tetrasulphonate (TPPS4) taken up by cells. Steady-state absorption and fluorescence spectra of TPPS4 in buffer solutions of different pH were used to identify a ring protonated species at pH less than or equal to 4. This dictation could also be distinguished from the unprotonated form by its altered fluorescence decay time (3.5 vs. 11.4 ns). In addition, time-resolved spectroscopy gave some evidence of a monocationic species existing at pH 6-9. This was concluded from the occurrence of another component with a decay time of 5 ns. Measurements of the spectral and kinetic properties of the fluorescence emission of single epithelial cells (RR1022) incubated with TPPS4 indicated that the sensitizer was mainly localized in a microenvironment with a pH of 5, a value which occurs intracellularly only within lysosomes. Cells kept in the dark exhibited the characteristic spectra of both the dication and the neutral form. The fluorescence decay showed two components with decay times of 2.6 ns and 10.6 ns. Irradiation of the cells changed the decay times to 4.6 ns and 13.4 ns and the dication fluorescence emission peak vanished, which is in accordance with the results obtained from buffer solutions at pH greater than or equal to 6. Therefore, we deduce that the photodynamic action leads to a rupture of the lysosomes and that the sensitizer is released into the surrounding cytoplasm.

Animals↗
PubMed1991

Photodynamic therapy of bladder cancer--uptake and phototoxicity of photosan in vitro.

The uptake of photosan and the intracellular sites of photoradiation-induced damage were investigated in vitro in bladder carcinoma cells and in normal bladder cells. Cells were examined by phase contrast, fluorescence and electron microscopy. The concentration of photosan, measured in microgram/10(6) cells, showed a good correlation to the incubation time. At all incubation times, control cells showed a lower uptake when compared with tumor cells. Following photodynamic therapy (PDT), phase-contrast microscopy revealed marked changes in tumor cells, whereas only minor effects could be detected at the cell membrane of the control cells. Following PDT, most of the investigated cells showed changes of the mitochondria and cytoplasma. These changes consisted of dissolution of the cristae, predominantly in the central part of the mitochondria. Twenty-four hours after PDT the shape of the mitochondria had changed markedly and the cristae were found to be completely destroyed. Moreover, the cystoplasma showed numerous vacuoles, and the number of mitochondria was decreased compared to non-treated cells.

Antineoplastic Agents↗
PubMed1990

Competition between photobleaching and fluorescence increase of photosensitizing porphyrins and tetrasulphonated chloroaluminiumphthalocyanine.

The time-dependent fluorescence changes of photosensitizing porphyrins and tetrasulphonated chloroaluminiumphthalocyanine (A1C1SPc) were measured at different intracellular sites using video-enhanced microscopy and image processing. To obtain variations in intracellular fluorescence intensity, different radiant exposures of a Kr+ laser-pumped dye laser were delivered via a 600 microns plastic-clad silica fibre connected to the microscope. During irradiation, competition between photobleaching and fluorescence increase of the different dyes was observed. The porphyrins normally showed photobleaching, which was dependent on the sensitizer and its specific accumulation within the cell. Photobleaching was less pronounced for hydrophilic uroporphyrin than for more hydrophobic dyes. In contrast with an almost exponential decrease in porphyrin fluorescence with increasing light dose, the fluorescence intensity of A1C1SPc significantly increased at the beginning of irradiation, and could be correlated with intracellular deaggregation.

Animals↗
PubMed1989

Intracellular localization of photosensitizers.

The intracellular localization of photosensitizers can be studied by different methods. One method involves homogenization of the cells followed by differential ultracentrifugation which leads to fractions enriched in nuclear, mitochondrial, and microsomal material as well as a supernatant fraction. More detailed information can be obtained by electron microscopy of cells exposed to light in the presence of photosensitizers. This method is based on the assumption that damage is primarily induced at intracellular sites where the concentration of photosensitizer is high. By irradiating the cells at 6 degrees C, where biochemical reactions are slow, and then incubating them for different times at 37 degrees C, it is possible to follow the development of damage. The amount of photosensitized damage to enzymes or cell functions whose localization in the cells is known gives information about the intracellular localization of the sensitizer. Fluorescence microscopy is the most direct method and is widely applicable because most photosensitizers fluoresce. Lipophilic dyes generally localize in membrane structures. In future more attention should be paid to the localization of dyes in lysosomes, as suggested by early reports. Mitochondria, the endoplasmic reticulum and nuclear membrane are other important loci for intracellular localization of sensitizers.

Animals↗
PubMed1988

Intracellular distribution of photosensitizing porphyrins measured by video-enhanced fluorescence microscopy.

The intracellular distribution of photosensitizing porphyrins with different amounts of aggregates was measured in a fluorescence microscope using video-intensified detection and image processing. Porphyrin fluorescence was localized in the plasma membrane, cytoplasm, nuclear membrane and nucleoli. The fluorescence intensity in the plasma membrane decreased with increasing aggregation of the photosensitizers. In addition, a redistribution of the porphyrin molecules from the plasma membrane to the nuclear membrane and adjacent intracellular sites was observed with increasing incubation time. Photobleaching of porphyrin fluorescence was most pronounced in the plasma membrane and least efficient within the nucleoli.

Chromatography, High Pressure Liquid↗
PubMedDate not supplied

[Fluorescence spectroscopy in selective percutaneous nucleotomy using the excimer laser--experimental studies].

Our intention was to test fluorescence spectroscopy as a possibility for selective laser ablation. In an experimental setup a XeCl excimer laser (308 nm) with pulse energies between 5 and 50 mJ was used for fluorescence excitation in 20 human and 100 pig vertebral segments. Tissue fluorescence was detected via a quartz fiber bundle and analyzed by a polychromator and optical multichannel analyzer. Both low and high energy levels led to a broad band fluorescence of nucleus pulposus, intervertebral muscle, spinal cord and vertebral end plate and a typical fluorescence of anulus fibrosus with peak maxima at 385 and 435 nm. Tissue fluorescence was independent of surrounding medium (with air and normal saline used) and excitation energy. Borderlines between different tissues could be clearly indicated in all cases. Using intensity rations at 350, 385 and 435 nm (I1 = I350/I385, I2 = I385/I435) tissue discrimination was possible in all cases. This results demonstrate, that discrimination between intervertebral disc and surrounding tissue is possible and could be used for selective percutaneous laser nucleotomy in the near future.

Animals↗