Visualisation of mitochondria in living neurons with single- and two-photon fluorescence laser microscopy

In this work we investigated the relative merits of conventional single-photon confocal laser scanning fluorescence microscopy (CLSM) and two-photon laser scanning fluorescence microscopy (2p-LSM) for the study of mitochondria in living neurons. Dorsal root ganglion neurons were loaded with the mitochondrion-specific fluorescent dye JC-1, the ratio between red (J-aggregates) and green (monomer) fluorescence of which reflects mitochondrial membrane potential. Cells were illuminated at 488 nm for single-photon excitation or at 870 nm for two-photon excitation. In both modalities we found that mitochondria showed: (i) similar appearance; (ii) similar fluorescence ratio values over both whole cell bodies and individual mitochondria; and (iii) similar responses to mitochondrial uncoupler, which dropped the ratio values by 50%. However, 2p-LSM exhibited several advantages over CLSM: (i) better signal/noise ratio in the green emission channel; (ii) less phototoxicity upon repetitive scanning in the focal plane; and (iii) no significant loss of image quality upon repetitive scans in the z direction. We conclude that, while both techniques enable visualisation of individual mitochondria in living cells, 2p-LSM has significant advantages for physiological work requiring time-lapse experiments or four-dimensional reconstructions of mitochondria.     

Computer-generated, three-dimensional reconstruction of histological parallel serial sections displaying microvascular and glandular structures in human endometrium

This paper describes a technique to develop high-resolution three-dimensional (3D) images of microvasculature structures in curettage, hysterectomy or endometrial resection biopsies using parallel histological serial sections. Employing a labelled streptavidin-biotin-alkaline phosphatase (LSAB(+)) method and visualising by using DAB(+) with the primary antibody, mouse anti human Q-Bend-10, the images were directly digitised from a light microscope into the KS400 Universal Image Processing and Analysis software via a CCD colour camera; binary images of the structures were created and the binary images were exported into VoxBlast 3D rendering software to view still and rotating 3D images on a computer monitor. This in turn enabled hard copies of the full sequence to be printed.     

Origin of the high piezoelectric response in PbZr1-xTixO3

High resolution x-ray powder diffraction measurements on poled PbZr1-xTixO3 (PZT) ceramic samples close to the rhombohedral-tetragonal phase boundary (the so-called morphotropic phase boundary) have shown that for both rhombohedral and tetragonal compositions the piezoelectric elongation of the unit cell does not occur along the polar directions but along those directions associated with the monoclinic distortion. This work provides the first direct evidence for the origin of the very high piezoelectricity in PZT.     

Eliminating false positive C4 sugar tests on New Zealand Manuka honey

Carbon isotope analyses (δ¹³C) of some New Zealand Manuka honeys show that they often fail the internationally recognised Association of Official Analytical Chemists sugar test (AOAC method 998.12) which detects added C₄ sugar, although these honeys are from unadulterated sources. Failure of these high value products is detrimental to the New Zealand honey industry, not only in lost export revenue, but also in brand and market reputation damage. The standard AOAC test compares the carbon isotope value of the whole honey and corresponding protein isolated from the same honey. Differences between whole honey and protein δ¹³C values should not be greater than +1.0‰, as it indicates the possibility of adulteration with syrups or sugars from C₄ plants such as high fructose corn syrup or cane sugar. We have determined that during the standard AOAC method, pollen and other insoluble components are isolated with the flocculated protein. These non‐protein components have isotope values which are considerably different from those of the pure protein, and can shift the apparent δ¹³C value of protein further away from the δ¹³C value of the whole honey, giving a false positive result for added C₄ sugar. To eliminate a false positive C₄ sugar test for Manuka honey, prior removal of pollen and other insoluble material from the honey is necessary to ensure that only the pure protein is isolated. This will enable a true comparison between whole honey and protein δ¹³C isotopes. Furthermore, we strongly suggest this modification to the AOAC method be universally adopted for all honey C₄ sugar tests. Copyright © 2010 John Wiley & Sons, Ltd.     

Large two-dimensional clusters of equal-area bubbles: The influence of the boundary in determining the minimum-energy configuration

Finite monodisperse two-dimensional clusters of bubbles are shown to behave like a crystal rather than a liquid. They attain their lowest-energy configuration when they find an arrangement close to a hexagonal lattice, as predicted by the Wulff construction. The deviation from hexagonal shape of the bubbles in a cluster and the variation in bubble pressure throughout the structure are calculated. It is shown that the effect of reducing the length of the boundary is negligible, explaining why configurations retaining the structure of the lattice will give the lowest energy.     

The rate at which energy decays in a damped String

The energy in a string subject to positive viscous damping is known to decay exponentially in time. Under the assumption that the damping is of bounded variation, we identify the best rate of decay with the supremum of the real part of the spectrum of the infinitesimal generator of the underlying semigroup. We analyze the spectrum of this nonselfadjoint operator in some detail. Our bounds on its real eigenvalues and asymptotic form of its large eigenvalues translate into criteria for over/underdamping and a proof that the decay rate achieves its (negative) minimum over those dampings whose total variation does not exceed a prescribed value.     

Oxygen diffusion in poly(dimethyl siloxane) using fluorescence quenching. II. Filled samples

The method of fluorescence quenching has been used to measure the transport properties of oxygen in poly(dimethyl siloxane) (PDMS) filled with small weight fractions of fumed silica. The experimental results show that the diffusion coefficient of oxygen and the pre-exponential term in the Arrhenius relationship are reduced, but the activation energy is not affected. In the pressure and temperature ranges used in these experiments, the diffusion coefficient was found to be independent of both the oxygen and fluorophor concentrations. A discussion concerning the effect of fillers on the pre-exponential term is presented.     

A Test of Multivariate Independence Based on a Single Factor Model

A test of the independence of two sets of variables is developed to have high power against a special family of dependence. In this each set of variables has the structure of a single factor model and the dependence is solely via the correlation γ between the underlying latent variables. This is a model with only one nonzero canonical correlation. It is shown that a test based on the maximum likelihood estimate of γ is appreciably more powerful than that based on r₁, the largest sample canonical correlation. If, however, the model is used, not just as a family of alternatives but as the basis for interpretation, and if substantial cross-correlation is present then the procedure is essentially equivalent to the use of r₁.