Component-resolved bleaching spectra of quartz optically stimulated luminescence: preliminary results and implications for dating

Bleaching spectra of the ‘fast’ and ‘medium’ optically stimulated luminescence (OSL) components of quartz are reported. A dependence of photoionization cross-section, σ, on wavelength was observed for the fast and medium components and a significant difference in their responses to stimulation wavelength was found. The ratio of the fast and medium photoionization cross-sections, σfast/σmedium, varied from 30.6 when stimulated with View the MathML source light to 1.4 at View the MathML source. At View the MathML source the fast and medium photoionization cross-sections were found to be sufficiently different that infrared bleaching at raised temperatures allowed the selective removal of the fast component with negligible depletion of the medium. A method for optically separating the OSL components of quartz is suggested, based on the wavelength dependence of photoionization cross-sections.

Viral nucleolar localisation signals determine dynamic trafficking within the nucleolus

Localisation of both viral and cellular proteins to the nucleolus is determined by a variety of factors including nucleolar localisation signals (NoLSs), but how these signals operate is not clearly understood. The nucleolar trafficking of wild type viral proteins and chimeric proteins, which contain altered NoLSs, were compared to investigate the role of NoLSs in dynamic nucleolar trafficking. Three viral proteins from diverse viruses were selected which localised to the nucleolus; the coronavirus infectious bronchitis virus nucleocapsid (N) protein, the herpesvirus saimiri ORF57 protein and the HIV-1 Rev protein. The chimeric proteins were N protein and ORF57 protein which had their own NoLS replaced with those from ORF57 and Rev proteins, respectively. By analysing the sub-cellular localisation and trafficking of these viral proteins and their chimeras within and between nucleoli using confocal microscopy and photo-bleaching we show that NoLSs are responsible for different nucleolar localisations and trafficking rates.

Monitoring growth of hard corals as performance indicators for coral reefs

Digital videophotography, computer image analysis and physical measurements have been used to monitor sedimentation rates, coral cover, genera richness, rugosity and estimated recruitment dates of massive corals at three different sites in the Wakatobi Marine National Park, Indonesia, and on the reefs around Discovery Bay, Jamaica. Semi-structured interviews with key stakeholders in the Wakatobi Marine National Park indicated that coral mining was extensively practised, and is responsible for the absence of large non-branching corals on the Sampela reef Blast fishing is also practised in the Wakatobi Marine Park, and the authors, together with students, showed that blast fishing resulted in coral bleaching and not mortality of two Porites lutea colonies. In addition, we showed that monitoring of bleaching in Porites colonies induced by blast fishing could be a useful way of monitoring blast fishing practices in susceptible areas in the Indo-Pacific. The techniques used in this study are appropriate for use by volunteers with sufficient training, and provide excellent projects for dissertation students reading undergraduate degrees.

A multi-level analytical approach for detection and visualization of intracellular NO production and nitrosation events using diaminofluoresceins

Diaminofluoresceins are widely used probes for detection and intracellular localization of NO formation in cultured/isolated cells and intact tissues. The fluorinated derivative, 4-amino-5-methylamino-2′,7′-difluorofluorescein (DAF-FM), has gained increasing popularity in recent years due to its improved NO-sensitivity, pH-stability, and resistance to photo-bleaching compared to the first-generation compound, DAF-2. Detection of NO production by either reagent relies on conversion of the parent compound into a fluorescent triazole, DAF-FM-T and DAF-2-T, respectively. While this reaction is specific for NO and/or reactive nitrosating species, it is also affected by the presence of oxidants/antioxidants. Moreover, the reaction with other molecules can lead to the formation of fluorescent products other than the expected triazole. Thus additional controls and structural confirmation of the reaction products are essential. Using human red blood cells as an exemplary cellular system we here describe robust protocols for the analysis of intracellular DAF-FM-T formation using an array of fluorescence-based methods (laser-scanning fluorescence microscopy, flow cytometry and fluorimetry) and analytical separation techniques (reversed-phase HPLC and LC-MS/MS). When used in combination, these assays afford unequivocal identification of the fluorescent signal as being derived from NO and are applicable to most other cellular systems without or with only minor modifications.

Assessing the completeness of optical resetting of quartz OSL in the natural environment

Resetting of previously accumulated optically stimulated luminescence (OSL) signals during transport of sediment is a fundamental requirement for reliable optical dating. The completeness of optical resetting of 46 modern-age quartz samples from a variety of depositional environments was examined. All equivalent dose (De) estimates were View the MathML source, with the majority of aeolian samples View the MathML source, and fluvial samples View the MathML source. The OSL signal of quartz originates from several trap types with different rates of charge loss during illumination. As such, incomplete bleaching may be identifiable as an increase in De from easy-to-bleach through to hard-to-bleach components. For all modern fluvial samples with non-zero De values, SAR De(t) analysis and component-resolved linearly modulated OSL (LM OSL) De estimates showed this to be the case, implying incomplete resetting of previously accumulated charge. LM OSL measurements were also made to investigate the extent of bleaching of the slow components in the natural environment. In aeolian sediments examined, the natural LM OSL was effectively zero (i.e. all components were fully reset). The slow components of modern fluvial samples displayed measurable residual signals up to 15 Gy.

Identification of partial resetting using De as a function of illumination time

Modern age samples from various depositional environments were examined for signal resetting. For 19 modern aeolian/beach samples all De values obtained were View the MathML source, with ∼70% having View the MathML source. For 21 fluvial/colluvial samples, all De values were View the MathML source with ∼80% being View the MathML source. De as a function of illumination (OSL measurement) time (De(t)) plots were examined for all samples. Based on previous laboratory experiments, increases in De(t) were expected for partially reset samples, and constant De(t) for fully reset samples. All aeolian samples, both modern age and additional ‘young’ samples (<1000 years), showed constant (flat) De(t) while all modern, non-zero De, fluvial/colluvial samples showed increasing De(t). ‘Replacement plots’, where a regenerated signal is substituted for the natural, yielded constant (flat) De(t). These findings support strongly the use of De(t) as a method of identifying incomplete resetting in fluvial samples. Potential complicating factors, such as illumination (bleaching) spectrum, thermal instability and component composition are discussed and a series of internal checks on the applicability of the De(t) for each individual aliquot/grain level are outlined.

Further investigations of the quartz optically stimulated luminescence components using linear modulation

The optically stimulated luminescence (OSL) from quartz is known to be the sum of several components with different rates of charge loss, originating from different trap types. The OSL components are clearly distinguished using the linear modulation (LM OSL) technique. A variety of pre-treatment and measurement conditions have been used on sedimentary samples in conjunction with linearly modulated optical stimulation to study in detail the behaviour of the OSL components of quartz. Single aliquots of different quartz samples have been found to contain typically five or six common LM OSL components when stimulated at View the MathML source. The components have been parameterised in terms of thermal stability (i.e. E and s), photoionisation cross-section energy dependence and dose response. The results of studies concerning applications of component-resolved LM OSL measurements on quartz are also presented. These include the detection of partial bleaching in young samples, use of ‘stepped wavelength’ stimulation to observe OSL from single components and attempts to extend the age range of quartz OSL dating.