ALBERT

Notes: Abstract Confocal fluorescence microscopy enables visualisation and quantitation of fluorescent probes at high resolution deep within intact tissues, with minimal disturbance both of cell–cell interactions and the mechanical, ionic and physiological effects of the extracellular matrix. We illustrate the principles of multiple-parameter 3-D (x,y,z) imaging using reconstruction of nuclear channels in mammalian cells. Repeated sampling in time generates 4-D (x,y,z,t) images which can be used to follow dynamic changes, such as blue-light-dependent chloroplast re-orientation, in intact tissues. Quantitative measurements from multi-dimensional images require calibration of the spatial dimensions of the image and the fluorescence intensity response. This must be determined throughout the volume, which must be sampled to correct for geometric distortion as well as photometric errors arising from the complete optical system, including the specimen. The effects of specimen calibration are illustrated for morphological analysis of stomatal closing responses to abscisic acid in Commelina from 4-D images. Calibrated 4-D imaging allows direct volume measurements and we have followed volume regulation of chondrocytes in cartilage explants during osmotic perturbation. In intact cartilage, unlike in isolated cells, the chondrocytes exhibit volume regulatory mechanisms. In other cases, the fluorescence intensity of the probe may be related to a physiological parameter of interest and changes in its distribution within the cell. Optical sectioning permits discrimination of signal in separate compartments within the cell and can be used to follow transport events between different organelles. We illustrate 3-D (x,y,t) measurements of vacuolar glutathione conjugate pump activity in intact roots of Arabidopsis by following the sequestration of a fluorescent conjugate between glutathione and monochlorobimane. Dynamic measurements of protein localisation are now possible following the introduction of chimeric fusion proteins with green fluorescent protein (GFP) from Aequoria victoria. We have analysed the disposition of heterochromatin in nuclei of living Schizosaccharomyces pombe cells expressing a chimeric construct between Swi6 and GFP. Heterochromatin dynamics can be followed throughout mitosis in 4-D (x,y,z,t) images. Statistical analysis of the fluorescence histograms from each nucleus over time provides quantitative support for aggregation and dispersion of Swi6-GFP clusters during mitosis, rather than dissociation of Swi6 from the heterochromatin. A wide range of single-wavelength and ratio probes are available for imaging different ion activities. We compare 3-D (x,y,t) measurements of ion activities made using single-wavelength (Fluo-3 for calcium) and ratio (BCECF for pH) measurements, using stomatal responses in Vicia faba to peptides from the auxin-binding protein of maize and tip growth in pollen tubes of Lilium longiflorum as examples. Ratioing techniques have many advantages for quantitative fluorescence measurements and we conclude with a discussion of techniques to develop ratioing of single-wavelength probes against alternative references, such as DNA, protein or cell wall material.

Notes: Crystalline SiGe alloy sheet polymers were topotactically prepared from epitaxially grown calcium germanosilicide Ca(Si1−xGex)2 precursor films in the whole composition range. These polygermanosilynes are found to be a well-defined mixture of the known siloxene and polygermyne sheet polymers with the OH groups exclusively bonded to silicon. The optical properties determined by photoluminescence and optical reflection measurements identify the mixed SiGe sheet polymers as direct semiconductors with efficient luminescence tunable in the energy range between 2.4 and 1.3 eV. © 2001 American Institute of Physics.

Notes: The level of glutathione (GSH) in plants is important in defence reactions against biotic and abiotic stresses and can place considerable demand of the sulphur assimilation pathway. Enzymes involved in sulphur assimilation and GSH metabolism are not evenly distributed between different subcellular compartments or between different cell types in leaves or roots; however, there is little information on the effect that such asymmetries have on the actual GSH concentration in each compartment or cell type. In the present study in situ labelling with monochlorobimane (MCB) in combination with confocal laser scanning microscopy was used to quantify GSH in each of the main cell types of poplar leaves from fluorescence of the GSB conjugate formed. Comparison of results from the in situ approach with total GSH levels measured in vitro by high-performance liquid chromatography suggested that only the cytosolic GSH pool was labelled using this approach. This suggests that an appropriate GST was not present within the chloroplasts to catalyse the conjugation reaction and that chloroplastic GSH does not rapidly exchange with the cytoplasmic pool under the conditions of the assay. Cytosolic GSH levels were between 0.2 and 0.3 mm for both photosynthetic and non-photosynthetic (epidermal) cell types in wild-type poplar leaves. Cytosolic levels increased by around two-fold in transgenic poplars over-expressing bacterial γ-glutamylsynthetase (γ-ECS) in the cytosol of all cell types, but there was no concomitant increase in the chloroplastic GSH pool.

Notes: "REVIEWS"

Notes: BOOK REVIEWS

Notes: A method of measuring the difference between the lattice parameter of a region of an unknown sample and that of a region of a standard reference crystal to a sensitivity of 1 part in 109 is presented. Problems inherent in multiple-beam arrangements due to sample strains and non-uniformity have been overcome by the use of a new double-source arrangement in which the two X-ray beams sample the same spot on a crystal under study. Ways of identifying and preventing errors from significant mechanical and thermal effects arising in the sensitivity region explored are indicated.

Notes: Zusammenfassung Die thermal bedingte Entwicklung permischer Schichten in der »Breccia Pipes« Region der Colorado-Hochebene wurde mittels dreier Geothermometer rekonstruiert. Die Rekonstruktion der Ablagerungsgeschichte deutet auf maximale Temperaturen zwischen 100 ° und 140 °C hin. Eine Spaltspuren-Analyse erbrachte als Ergebnis eine maximale Temperatur von 137 °C, die über 50 Ma. integriert wurde. Diese Ergebnisse stimmen mit der Minimum-Temperatur überein, die mit Hilfe von Flüssigkeitseinschlüssen (75°–140 °C) errechnet wurden. Die Hypothesen der Spaltspuren-Analyse und der Ablagerungsgeschichte wurden in ein kinetisches Modell der Reifung von organischen Substanzen miteinbezogen. Die errechneten Durchschnitts-temperaturen liegen bei 445° und 421 °C. Ein Vergleich mit Daten der organischen Geochemie aus der permischen »Toroweap« Formation zeigt, daß die zwei oben genannten Hypothesen es erlauben, thermale Rahmenbedingungen für die Entwicklung organischer Substanzen festzusetzen. Die wichtigsten Schlüsse, die aus dieser Studie gezogen werden können sind das Alter des Beginns der Ölbildungsphase und der regionalen Hebungsphase (Laramid-Orogenese) sowie die Spannweite der maximalen Temperaturen.

Notes: Abstract The ongoing Chase Delphi study of fusion power has employed a panel of over 100 experts from 13 countries. The use of fusion for synfuel production was emphasized. The panelists believed that the length of time to successful demonstrations and commercialization of fusion could be substantially reduced over current estimates given optimal funding, and that a hydrocarbon producing reactor is feasible with minimal adverse impact on the environment.