No abstract

Phosphorylation of the chromatin protein H2AX (forming γH2AX) is implicated in the repair of DNA double strand breaks (DSB's); a large number of H2AX molecules become phosphorylated at the sites of DSB's. Fluorescent staining of the cell nuclei for γH2AX, via an antibody, visualises the formation of these foci, allowing the quantification of DNA DSB's and forming the basis for a sensitive biological dosimeter of ionising radiation.We describe an automated fluorescence microscopy system, including automated image processing, to count γH2AX foci. The image processing is performed by a Hough transform based algorithm, CHARM, which has wide applicability for the detection and analysis of cells and cell colonies. This algorithm and its applications for cell nucleus and foci detection will be described. The system also relies heavily on robust control software, written using multi-threaded cbased modules in LabWindows/CVI that adapt to the timing requirements of a particular experiment for optimised slide/plate scanning and mosaicing, making use of modern multi-core processors. The system forms the basis of a general purpose high-content screening platform with wide ranging applications in live and fixed cell imaging and tissue micro arrays, that in future, can incorporate spectrally and time-resolved information.

Spontaneous rotational Raman scattering spectroscopy is used to acquire measurements of the mean and root mean square (rms) temperature fluctuations in turbulent, high-velocity heated jets. Raman spectra in air were obtained across a matrix of radial and axial locations downstream from a 50 mm diameter nozzle operating from subsonic to supersonic conditions over a wide range of temperatures and Mach numbers, in accordance with the Tanna matrix frequently used in jet noise studies. These data were acquired in the hostile, high noise (115 dB) environment of a large scale open air test facility at NASA Glenn Research Center (GRC). Temperature estimates were determined by performing non-linear least squares fitting of the single shot spectra to the theoretical rotational Stokes spectra of N2 and O2. The laser employed in this study was a high energy, long-pulsed, frequency doubled Nd:YAG laser. One thousand single-shot spectra were acquired at each spatial coordinate. Mean temperature and rms temperature variations were calculated at each measurement location. Excellent agreement between the averaged and single-shot temperatures was observed with an accuracy better than 2.5% for temperature, and rms variations in temperature between  ±2.2% at 296 K and  ±4.5% at 850 K. The mean and normalized rms temperatures measured here were then compared to NASA’s Consensus data set of PIV velocity and turbulence measurements in similar jet flows. The results of this and planned follow-on studies will support NASA GRC’s development of physics-based jet noise prediction, turbulence modeling and aeroacoustic source modeling codes.

The time-resolved emission spectra of a series of di-/3-substituted 1 .«-dinaphthylalkanes have been investigated in isooctane at room temperature. Phosphorescence from the triplet excimer has been observed for the longer alkyl chain derivatives ( = 3, 4) but not from the short-chain species ( = 1, 2). Under identical experimental conditions, the excimer phosphorescence from the ß derivatives is blue shifted and is much weaker than that of the corresponding a isomers. Furthermore, the decay rates are much greater for the di-ß molecules. These results confirm the previously proposed "L"-shaped structure of the triplet excimers of naphthalene.