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At least 613 records · Page 34Linked to original sources
PubMed1997

[Fluorescence lifetimes of squamous carcinoma, adenocarcinoma and normal human lung tissues in vitro by time-resolved fluorescence].

Using pulsed dye laser 632.8nm, lifetimes of adenocarcinoma, squamous carcinoma and normal human lung tissues in vitro have been measured by time-resolved fluorescence as 5.1, 5.6 and 6.3 ns respectively. Time-resolved fluorescence shows that the decay time of tumor tissues is faster than that of normal tissues. The results may offer new diagnostic method to determine cancer.

Adenocarcinoma↗
PubMed1986

[Demonstration of the intracellular accumulation of intermediary fluorescent metabolites of various polycyclic aromatic hydrocarbons by resolution of the fluorescence spectra of isolated living cells].

Using microspectrofluorimetry, we have studied some spectral modifications induced by the accumulation of fluorescent metabolites from some PAH in single living cells (3T3 and RTG2 fibroblasts). Whatever the parent compound under study, no spectral deformation has been detected in the 3T3 cells. On the contrary, the characteristic fluorescence spectra were strongly modified in RTG2 cells when the parent compound was either B(a)P or CPAP. The B(a)P metabolites have been identified as 3-OH-B(a)P and 9-OH-B(a)P.

Animals↗
PubMed1978

[The differentiation of clostridial microcultures on nuclepore-filters by fluorescent antibodies as a rapid screening test for spores. II. Application of fluorescent antibodies (author's transl)].

The study deals with the question of whether a combination of microcolony-cultivation on polycarbonate-membrane-filters with the fluorescent-antibody-method will lead to a rapid-diagnosis method for clostridial-spores. Formaldehyd-inactivated vegetative cells of eight different clostridial-strains (Table 1) were used for the immunisation of rabbits according to the method published by Böhm and Strauch (5). The results of the immunisation procedure, developed for aerobic-sporeformers, showed, that satisfactory results could not be obtained in all cases (Table 2). The eight conjugates were tested against microcolonies of 32 strains of different clostridial-species on membrane-filters, produced by the technique described in the first communication. The results showed (Table 4) that differentiation was always possible when homologous antisera were used. In most cases, in order to differentiate between the various species, conjugates of several serotypes had to be employed together. However due to their uniform antigenic nature, Cl. feseri and Cl. tetani could both be identified by using one strain for each to immunize. With clostridial-spores the species could be identified within a period of 6 to 12 hours by using the membrane-filter-fluorescent-antibody technique.

Antibodies, Bacterial↗
PubMed1980

Protein synthesis in rabbit reticulocytes. A study of the mechanism of interreaction of fluorescently labeled co-eIF-2A with eIF-2 using fluorescence polarization.

5-Dimethylaminonaphthalene-1-sulfonyl (dansyl)-Co-eIF-2A was prepared using homogeneous Co-eIF-2A. Dansyl-Co-eIF-2A was as active as untreated Co-eIF-2A when assayed for stimulation of ternary complex formation and also for protection of the ternary complex from dissociation by aurintricarboxylic acid. The mechanism of interaction of dansyl-Co-eIF-2A with eIF-2 was studied by measuring changes in fluorescence polarization. These studies indicate that dansyl-Co-eIF-2A interacts specifically with the ternary complex and does not interact with free eIF-2 or with two other high molecular weight protein complexes, Co-eIF-2B and Co-eIF-2C. Mg2+ inhibits ternary complex formation by eIF-2 and Co-eIF-2C relieves this Mg2+ inhibition of ternary complex formation. In both cases, the changes in fluorescence polarization of dansyl-Co-eIF-2A correlate well with the extent of ternary complex formed.

Animals↗
PubMed1998

Fluorescence in situ hybridization of progenitor cells obtained by fluorescence-activated cell sorting for the detection of cells affected by chromosome abnormality trisomy 8 in patients with myelodysplastic syndromes.

Myelodysplastic syndrome (MDS) is believed to be a stem-cell disorder involving cytopenia and dysplastic changes in three hematopoietic lineages. However, the involvement of pluripotent stem cells and progenitor cells has not been clarified conclusively. To address this issue, we used fluorescence in situ hybridization (FISH) of blood and bone marrow (BM) smears for mature cells and FISH of cells sorted by fluorescence-activated cell sorting for progenitor cells. Seven patients with MDS associated with trisomy 8 were studied. FISH showed +8 in granulocytes, monocytes, and erythroblasts, but not in lymphocytes. Sorted cells of T (CD3(+)), B (CD19(+)), and NK cells (CD3(-)CD56(+)) from peripheral blood did not contain +8, nor did CD34(+) subpopulations from BM including B (CD34(+)CD19(+)), T/NK (CD34(+)CD7(+)) progenitors, and pluripotent stem cells (CD34(+)Thy1(+)). The +8 chromosome abnormality was identified in stem cells only at the level of colony-forming unit of granulocyte-erythrocyte-macrophage-megakaryocyte (CFU-GEMM; CD34(+)CD33(+)). It may thus be concluded that cells affected by trisomy 8 in the context of MDS are at the CFU-GEMM level and that cells of lymphoid lineage are not involved. These results provide new insights into the biology of MDS and suggest that intensive chemotherapy and autologous BM transplantation may become important therapeutic strategies.

Adult↗
PubMed1998

Lineage involvement of stem cells bearing the philadelphia chromosome in chronic myeloid leukemia in the chronic phase as shown by a combination of fluorescence-activated cell sorting and fluorescence in situ hybridization.

Chronic myeloid leukemia (CML) is thought to arise from a pluripotent hematopoietic stem cell that has undergone a reciprocal translocation between the BCR gene on chromosome 22 and the ABL proto-oncogene on chromosome 9. This rearrangement results in a shortened chromosome 22, designated the Philadelphia (Ph) chromosome. The Ph chromosome has been found in cells from all hematopoietic lineages except mature T lymphocytes. To examine this issue, we combined fluorescence-activated cell sorting (FACS) and fluorescence in situ hybridization (FISH) to study lineage involvement of mature cells and stem cells in 12 patients with CML in the chronic phase. We found Ph chromosomes in myeloid cells and most B lymphocytes (CD19(+)) but not in mature T cells (CD3(+)) or natural killer (NK) cells (CD3(-)56(+)). Moreover, evidence of BCR/ABL fusion was found in pluripotent stem cells (CD34(+)Thy-1(+)), B-progenitor cells (CD34(+)CD19(+)), T/NK progenitor cells (CD34(+)CD7(+) cells), and T progenitor cells (CD34(+)CD7(+)CD5(+)) with a frequency equal to that in all CD34(+) cells isolated by FACS from bone marrow cells. T lymphocytes showed a marked decrease in Ph+ cells between progenitor cells and mature cells. Moreover, the ratios of Ph+ to Ph- cells in mature T cells and NK cells were below background levels, whereas Ph+ B lymphocytes also decreased during their maturation. These data suggest that Ph+ lymphocytes are eliminated during differentiation. In contrast to FISH of blood and bone marrow, which gives information principally about mature cells, the technique of "sorter FISH (FACS + FISH)" provides a powerful tool to explore the cytogenetic changes in immature cell populations of stem cell diseases based on immunophenotypes. Further clarification of genetic changes in stem cells could be achieved by using sorter FISH with monoclonal antibodies.

Adult↗
PubMed2005

Ultrafast fluorescence depolarisation in the yellow fluorescent protein due to its dimerisation.

Transient absorption spectroscopy with sub-100 fs time resolution was performed to investigate the oligomerisation behaviour of eYFP in solution. A single time constant tau(AD)=2.2+/-0.15 ps is sufficient to describe the time-resolved anisotropy decay up to at least 200 ps. The close contact of two protein barrels is deduced as the exclusive aggregation state in solution. From the final anisotropy r(infinity)=0.28+/-0.02, the underlying quaternary structure can be traced back to the somewhat distorted structure of the dimers of wt-GFP. The use of autofluorescent proteins as rulers in Förster resonance energy transfer (FRET) measurements may demand polarisation-sensitive detection of the fluorescence with high time resolution.

Bacterial Proteins↗
PubMed2004

Enhanced fluorescence of remote functionalized diaminodicyanoquinodimethanes in the solid state and fluorescence switching in a doped polymer by solvent vapors.

Remote functionalized zwitterionic diaminodicyanoquinodimethanes are found to exhibit a dramatic enhancement of light emission in the solid state and when doped in polymer films, as compared to the solution state. Crystal structure analysis of prototypical molecules reveals the role of the remote functionality in the solid state molecular organization. Semiempirical quantum chemical computations provide a viable model to explain the interesting phenomenon of fluorescence enhancement as arising from the inhibition of geometry relaxation of the vertical excited state to a nonemitting state. The reversible switching of a doped polymer film fluorescence triggered by solvent vapors is demonstrated.

Journal Article↗
PubMed2003

Laser-induced fluorescence with an OPO system. Part II: direct determination of lead content in seawater by electrothermal atomization-laser-excited atomic fluorescence (ETA-LEAF).

Fluorescence was induced by coupling a laser with an optical parametric oscillator (OPO) to develop an analytical method for the direct determination of lead content, at ultra-trace level, in seawater by electrothermal atomization-laser-excited atomic fluorescence (ETA-LEAF). The optimization of atomization conditions, laser pulse energy, and mainly temporal parameters allowed us to reach a 3 fg detection limit (0.3 ng L(-1)) despite the low repetition rate of the device. The expected error on predicted concentrations of lead, at trace levels, in seawater was below 15%.

Journal Article↗
PubMed2006

Early detection of plant stress from changes in distributions of chlorophyll a fluorescence parameters measured with fluorescence imaging.

In this work, we used barley leaves suffering from a stress, for measurements of chlorophyll a fluorescence with an imaging fluorometer. We compared selected fluorescence parameters (FP) determined from the measurements of control (no stress) and afterwards stressed sample by classical statistical comparison (Mann-Whitney test) and by statistical comparison of shapes of distributions of the FPs (two-sample Smirnov test). We have found that there exist examples where statistically significant difference is not revealed using the classical statistical comparison (for given critical level), but statistically significant difference is revealed using comparisons of distributions (for the same critical level). It implies that the shape of statistical distribution of a FP is more sensitive to a stress of a sample than median of the FP. Further, the comparison of changes in shapes of statistical distributions of FPs is therefore more suitable for early detection of plant stress than a classical statistical comparison. The observed changes in the distributions of FPs are discussed.

Journal Article↗
PubMed1986

Dibekacin assay in serum by automated fluorescence polarization immunoassay (Abbott Tdx): comparison with high-performance liquid chromatography, substrate-labelled fluorescent immunoassay and radioimmunoassay.

An immunoassay based on fluorescence polarization detection (FPIA) has been recently adapted for dibekacin. This has been compared with a reference method (high-performance liquid chromatography), and two other methods used in clinical laboratories for monitoring this aminoglycoside, namely substrate-labelled fluorescent immunoassay (SLFIA) and radioimmunoassay (RIA). FPIA was fast and more reliable than SLFIA or RIA, and offered therefore superior performance. However, its nominal cost per assay is high.

Journal Article↗
PubMed2006

Luminescence techniques and characterization of the morphology of polymer latices 2. Fluorescence lifetime, phosphorescence and fluorescence anisotropy studies.

Five poly(n-butyl methacrylate), PBMA, latex dispersions have been prepared, each incorporating a different fluorescent label, via a two-stage seeded emulsion polymerization. The resultant latices contain ca. 35% by weight total solids and are of 80 (+/-10) nm diameter as determined by photon correlation spectrometry. Luminescence spectroscopic techniques, namely fluorescence (and phosphorescence) excited state lifetime measurements in addition to time-resolved anisotropy experiments have provided useful information regarding the morphology, microviscosity and water permeability of the resultant particles. A picture of the PBMA colloid emerges of an interior which is highly viscous and water impermeable in nature. Indeed, the environment is protective enough to sustain room temperature stabilized phosphorescence from both an acenaphthylene and 9-phenanthrylmethyl methacrylate labeled dispersion through simple nitrogen purging of the solutions. However, the current spectroscopic measurements should be viewed with the knowledge that each luminescent label may fashion its own distinctive microenvironment within the latex during polymerization.

Journal Article↗
PubMed2005

X-ray absorption fine structure combined with x-ray fluorescence spectrometry. Part 15. Monitoring of vanadium site transformations on titania and in mesoporous titania by selective detection of the vanadium K alpha1 fluorescence.

X-ray absorption fine structure combined with X-ray fluorescence spectrometry was applied to various V+TiO2 hybrid samples. Emitted V K alpha1 fluorescence from the sample was selectively counted by using a high-energy-resolution (0.4 eV) spectrometer equipped with a Ge(331) crystal. Two advantages of this method, extremely high signal/background ratio and the compatibility of measurements in the atmosphere of reaction gas (in situ study in relation to heterogeneous catalysis), were effective at the V K-edge. Structure transformation of the V sites was spectroscopically followed for the V/TiO2 catalyst. The monooxo tetrahedral vanadate site was demonstrated to exist at 473 K. It transformed into dispersed species of 5-fold coordination in ambient air and further into polymeric VO(x) species in 0.85 kPa of water at 290 K. In the presence of 3.2 kPa of 2-propanol, dissociative adsorption of 2-propanol on the dispersed V species was strongly suggested at 290-473 K. In situ structure changes of V sites on TiO2 were reported by means of XAFS for the first time. The V(V) sites for the V/TiO2 catalysts were essentially identical with those for V supported on mesoporous (high-surface-area) TiO2 and V-TiO2 sample prepared by the sol-gel method. However, predominant V(IV) sites were found for mesoporous V-TiO2. The V(IV) sites substituted on the Ti sites of TiO2. When the molar ratio of V/Ti increased from 1/100 to 1/5.0, major octahedral V sites in the TiO2 matrix looked to transform into tetrahedral ones.

Journal Article↗
PubMed2006

Fluorescent organic nanoparticles of benzofuran-naphthyridine linked molecules: formation and fluorescence enhancement in aqueous media.

[structure: see text] Ethynyl-linked benzofuran-naphthyridine compounds show high-yield fluorescence with solvatochromic properties. One of the compounds, ABAN, has successfully formed fluorescent organic nanoparticles (FONs), for which the photophysical properties such as the spectral features and intensity are remarkably different from those at the molecular level (solution) and in bulk material. The results are tentatively rationalized by the FONs inducing coplanarization of the benzofuran-naphthyridine molecule to extend its effective conjugation length and hence increase the oscillator strength.

Journal Article↗
PubMed1997

Phloem Unloading in Sink Leaves of Nicotiana benthamiana: Comparison of a Fluorescent Solute with a Fluorescent Virus.

Using noninvasive imaging techniques, we compared phloem unloading of the membrane-impermeant, fluorescent solute carboxyfluorescein (CF) with that of potato virus X expressing the gene for the green fluorescent protein. Although systemic virus transport took considerably longer to occur than did CF transport, unloading of both solute and virus occurred predominantly from the class III vein network, a highly branched veinal system found between class II veins. The minor veins (classes IV and V) played no role in solute or virus import but were shown to be functional in xylem transport at the time of import by labeling with Texas Red dextran. After virus exit from the class III phloem, the minor veins eventually became infected by cell-to-cell virus movement from the mesophyll. During the sink/source transition, phloem unloading of CF was inhibited from class III veins before the cessation of phloem import through them, suggesting a symplastic isolation of the phloem in class III veins before its involvement in export. The progression of the sink/source transition for carbon was unaffected by the presence of the virus in the sink leaf. However, the virus was unable to cross the sink/source boundary for carbon that was present at the time of viral entry, suggesting a limited capacity for cell-to-cell virus movement into the apical (source) region of the leaf. A functional model of the sink/source transition in Nicotiana benthamiana is presented. This model provides a framework for the analysis of solute and virus movement in leaves.

Journal Article↗
PubMed2005

Characterization of sectioning fluorescence microscopy with thin uniform fluorescent layers: Sectioned Imaging Property or SIPcharts.

Thin, uniformly fluorescing reference layers can be used to characterize the imaging conditions in confocal, or more general, sectioning microscopy. Through-focus datasets of such layers obtained by standard microscope routines provide the basis for the approach. A set of parameters derived from these datasets is developed for defining a number of relevant sectioned imaging properties. The main characteristics of a particular imaging situation can then be summarized in a Sectioned Imaging Property-chart or SIPchart. We propose the use of such charts for the characterization of imaging properties in confocal and multiphoton microscopy. As such, they can be the basis for comparison of sectioned imaging condition characteristics, quality control, maintenance or reproduction of sectioned imaging conditions and other applications. Such charts could prove useful in documenting the more relevant properties of the instrumentation used in microscopy studies. The method carries the potential to provide the basis for a general characterization of sectioned imaging conditions as the layers employed can be characterized and fabricated to standard specifications. A limited number of such thin, uniformly fluorescing layers is available from our group for this purpose. Extension of the method to multiphoton microscopy is discussed.

Journal Article↗
PubMed1993

Inequivalence of fluorescent choline and ethanolamine phospholipids in the erythrocyte membrane: fluorescence lifetime determination in the frequency and time domain.

The fluorescence decay of 1-palmitoyl-2-diphenyl-hexatrienyl-propionyl-sn-glycero-3-phosphocholine and the respective ethanolamine derivative were determined in human erythrocyte ghost membranes by phase and modulation fluorometry. The obtained bimodal distribution decay model could independently be confirmed by pulse fluorometry and data analysis by the exponential series method. Distributional widths of labeled choline and ethanolamine phospholipids were different in ghost membranes but identical in phospholipid vesicles. Thus, it is concluded that both lipid classes containing the same fluorophore experience environments of different heterogeneity in the biological system.

Erythrocyte Membrane↗
PubMed1994

Time-resolved polarized fluorescence studies of the temperature adaptation in Bacillus subtilis using DPH and TMA-DPH fluorescent probes.

The validity of the concept of homeoviscous adaptation was tested for bacteria Bacillus subtilis. The Bacillus subtilis grown at 20 degrees C (referred to as Bs20) exhibit a considerable increase of branched anteiso-C15, the major fatty acid component of membrane lipids, relative to membranes grown at 40 degrees C (Bs40). The time-resolved fluorescence depolarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) and 1-[4-(trimethylamino)phenyl]-6-phenyl-1,3,5-hexatriene (TMA-DPH) showed that these changes in the lipid composition are accompanied by changes in a mean lipid order. In particular, the DPH order parameters and measured in Bs20 membranes at 18 degrees C and in Bs40 membranes at 45 degrees C, respectively, tend to be equal. This effect was less pronounced for TMA-DPH. Our observations suggest that a physical parallel to the changes of lipid composition is the maintenance of an optimal lipid order in the hydrophobic core of the cytoplasmic membranes. It can be interpreted as a tendency of Bacillus subtilis to keep the lateral pressure in its membranes at an optimal value, independent of the temperature of cultivation.

Adaptation, Physiological↗