Effect of hydrogen/air flow rates and scan rate on the flame ionisation detection response of phopholipids, and their quantitative and qualitative analysis by Iatroscan (Mark‐6s) TLC‐FID

The combined effect of scan speed, hydrogen and air flow rates on the flame ionization detection (FID) peak response of phospholipid classes has been studied to determine the optimum levels of these parameters. The phospholipid composition of different types of commercial lecithins, as well as lecithins combined with fish oils, has been analyzed by Iatroscan TLC‐FID Mark‐6s under optimized conditions. An air flow rate of 2 L/min, a hydrogen flow rate of 150–160 mL/min, and a scan speed of 30 s/rod seem to be the ideal conditions for scanning phospholipids with complete pyrolysis in the flame in the Mark‐6 model. Increasing the scan speed rapidly decreased the FID response. A hydrogen flow rate as high as 170 mL/min could be used at relatively low air flow rates (&#x003C2 L/min) and the response declined when both air flow rate and hydrogen flow rate increased simultaneously. Both linear and curvilinear relationships had highly significant correlations (p&#x003C0.01) with the sample load. Time course reactions, including the hydrolysis of phosphatidylserine using enzymes, can be successfully monitored by the Iatroscan TLC‐FID Chromarod system.

The male germ unit of Rhododendron : quantitative cytology, three-dimensional reconstruction, isolation and detection using fluorescent probes

The sperm cells of Rhododendron laetum and R. macgregoriae differentiate within the pollen tube about 24 h after germination in vitro. Threedimensional reconstruction shows that the sperm cells are paired together, and both have extensions that link with the tube nucleus, forming a male germ unit. Quantitative analysis shows that the sperm cells in each pair differ significantly in surface area, but not in cell volume nor in numbers of mitochondria or plastids. When isolated from pollen tubes by osmotic shock, the sperm cells became ellipsoidal and surrounded by their own plasma membrane, while a proportion remained in pairs linked by the inner tube plasma membrane. Both generative and sperm cells are visualized in pollen tube preparations by immunofluorescence with anti-tubulin and anti-actin monoclonal antibodies (MAbs) combined with H33258 fluorescence of the nuclei. Video-image processing shows the presence of an axial microtubule cage in the generative cells, and some microtubules are present in the cytoplasmic extensions that clasp the tube nucleus. Following sperm cell division, the extensive phragmoplast between the sperm nuclei is partitioned by the plasma membranes.

Qualitative and quantitative analysis of lipid classes in fish oils by thin-layer chromatography with an Iatroscan flame Ionization detector (TLC-FID) and liquid chromatography with an evaporative light scattering detector (LC-ELSD)

Combined effects of hydrogen and air flow rates on the peak response of selected neutral lipid classes (triacylglycerol, diacylglycerol, monoacylglycerol, free fatty acids, and ethyl esters) were studied to optimize and calibrate the Iatroscan Mk-6s Chromarod system for the qualitative and quantitative analysis of lipid classes by thin-layer chromatography (TLC) with flame ionization detection in fish oil during the transesterification process. Air flow rate of 2 L/min, hydrogen flow rate of 150-160 mL/min, and scan rate of 30 s/rod were found to be the optimum conditions. All samples were also analyzed by high performance liquid chromatography (HPLC) with evaporative light scattering detection. Quantitative results obtained by TLC with the flame ionization detection method were comparable to those obtained from HPLC with evaporative light scattering detection.

Fast colorimetric detection of copper ions using L-Cysteine functionalized gold nanoparticles

This communication reports an efficient visual detection method of Cu²⁺ by L-cysteine functionalized gold nanoparticles in aqueous solution. Upon exposure to Cu²⁺, the gold nanoparticle solution changed from red to blue, in response to surface plasmon absorption of dispersed and aggregated nanoparticles. This colorimetric sensor allows a rapid quantitative assay of Cu²⁺ down to the concentration range of 10⁻⁵ M. Recognition of Cu²⁺ and formation of the aggregates are proposed to occur via a 2 : 1 sandwich complex between L-cysteine and Cu²⁺.

Unification of the copper(I) binding affinities of the metallo-chaperones Atx1, Atox1, and related proteins : Detection probes and affinity standards

Literature estimates of metal-protein affinities are widely scattered for many systems, as highlighted by the class of metallo-chaperone proteins, which includes human Atox1. The discrepancies may be attributed to unreliable detection probes and/or inconsistent affinity standards. In this study, application of the four CuI ligand probes bicinchoninate, bathocuproine disulfonate, dithiothreitol (Dtt), and glutathione (GSH) is reviewed, and their CuI affinities are re-estimated and unified. Excess bicinchoninate or bathocuproine disulfonate reacts with CuI to yield distinct 1:2 chromatophoric complexes [CuIL2] 3- with formation constants β2 = 1017.2 and 1019.8 M-2, respectively. These constants do not depend on proton concentration for pH ≥7.0. Consequently, they are a pair of complementary and stable probes capable of detecting free Cu+ concentrations from 10-12 to 10-19 M. Dtt binds CuI with KD∼10-15 M at pH 7, but it is air-sensitive, and its CuI affinity varies with pH. The CuI binding properties of Atox1 and related proteins (including the fifth and sixth domains at the N terminus of the Wilson protein ATP7B) were assessed with these probes. The results demonstrate the following: (i) their use permits the stoichiometry of high affinity CuI binding and the individual quantitative affinities (KD values) to be determined reliably via noncompetitive and competitive reactions, respectively; (ii) the scattered literature values are unified by using reliable probes on a unified scale; and (iii) Atox1-type proteins bind CuI with sub-femtomolar affinities, consistent with tight control of labile Cu+ concentrations in living cells.

Corner Inlet and Nooramunga Habitat Mapping Project – Part 2 : High resolution mapping and change detection analysis using 2011 Rapid Eye Satellite Image compared to 2009 ALOS Satellite Image

Building on a habitat mapping project completed in 2011, Deakin University was commissioned by Parks Victoria (PV) to apply the same methodology and ground-truth data to a second, more recent and higher resolution satellite image to create habitat maps for areas within the Corner Inlet and Nooramunga Marine and Coastal Park and Ramsar area. A ground-truth data set using in situ video and still photographs was used to develop and assess predictive models of benthic marine habitat distributions incorporating data from both RapidEye satellite imagery (corrected for atmospheric and water column effects by CSIRO) and LiDAR (Light Detection and Ranging) bathymetry. This report describes the results of the mapping effort as well as the methodology used to produce these habitat maps.

Overall accuracies of habitat classifications were good, with error rates similar to or better than the earlier classification (>73 % and kappa values > 0.58 for both study localities). The RapidEye classification failed to accurately detect Pyura and reef habitat classes at the Corner Inlet locality, possibly due to differences in spectral frequencies. For comparison, these categories were combined into a ‘non-seagrass’ category, similar to the one used at the Nooramunga locality in the original classification. Habitats predicted with highest accuracies differed from the earlier classification and were Posidonia in Corner Inlet (89%), and bare sediment (no-visible seagrass class) in Nooramunga (90%). In the Corner Inlet locality reef and Pyura habitat categories were not distinguishable in the repeated classification and so were combined with bare sediments. The majority of remaining classification errors were due to the misclassification of Zosteraceae as bare sediment and vice versa. Dominant habitats were the same as those from the 2011 classification with some differences in extent. For the Corner Inlet study locality the no-visible seagrass category remained the most extensive (9059 ha), followed by Posidonia (5,513 ha) and Zosteraceae (5,504 ha). In Nooramunga no-visible seagrass (6,294 ha), Zosteraceae (3,122 ha) and wet saltmarsh (1,562 ha) habitat classes were most dominant.

Change detection analyses between the 2009 and 2011 imagery were undertaken as part of this project, following the analyses presented in Monk et al. (2011) and incorporating error estimates from both classifications. These analyses indicated some shifts in classification between Posidonia and Zosteraceae as well as a general reduction in the area of Zosteraceae. Issues with classification of mixed beds were apparent, particularly in the main Posidonia bed at Nooramunga where a mosaic of Zosteraceae and Posidonia was seen that was not evident in the ALOS classification. Results of a reanalysis of the 1998-2009 change detection illustrating effects of binning of mixed beds is also provided as an appendix.

This work has been successful in providing baseline maps at an improved level of detail using a repeatable method meaning that any future changes in intertidal and shallow water marine habitats may be assessed in a consistent way with quantitative error assessments. In wider use, these maps should also allow improved conservation planning, advance fisheries and catchment management, and progress infrastructure planning to limit impacts on the Inlet environment.

Quantitative secondary electron imaging for work function extraction at atomic level and layer identification of graphene

Two-dimensional (2D) materials usually have a layer-dependent work function, which require fast and accurate detection for the evaluation of their device performance. A detection technique with high throughput and high spatial resolution has not yet been explored. Using a scanning electron microscope, we have developed and implemented a quantitative analytical technique which allows effective extraction of the work function of graphene. This technique uses the secondary electron contrast and has nanometre-resolved layer information. The measurement of few-layer graphene flakes shows the variation of work function between graphene layers with a precision of less than 10 meV. It is expected that this technique will prove extremely useful for researchers in a broad range of fields due to its revolutionary throughput and accuracy.