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Biomedical subjects

Philip Heraud

Publications and source records attributed to Philip Heraud.

5 recordsLinked to original sources

Focal plane array infrared imaging: a new way to analyse leaf tissue.

* Here, a new approach to macromolecular imaging of leaf tissue using a multichannel focal plane array (FPA) infrared detector was compared with the proven method of infrared mapping with a synchrotron source, using transverse sections of leaves from a species of Eucalyptus. * A new histological method was developed, ideally suited to infrared spectroscopic analysis of leaf tissue. Spatial resolution and the signal-to-noise ratio of the FPA imaging and synchrotron mapping methods were compared. * An area of tissue 350 microm(2) required approx. 8 h to map using the synchrotron technique and approx. 2 min to image using the FPA. The two methods produced similar infrared images, which differentiated all tissue types in the leaves according to their macromolecular chemistry. * The synchrotron and FPA methods produced similar results, with the synchrotron method having superior signal-to-noise ratio and potentially better spatial resolution, whereas the FPA method had the advantage in terms of data acquisition time, expense and ease of use. FPA imaging offers a convenient, laboratory-based approach to microscopic chemical imaging of leaves.

Eucalyptus↗

Effects of nitrogen source and UV radiation on the growth, chlorophyll fluorescence and fatty acid composition of Phaeodactylum tricornutum and Chaetoceros muelleri (Bacillariophyceae).

Cultures of the marine diatoms Phaeodactylum tricornutum and Chaetoceros muelleri were grown in f/2 medium supplied with either nitrate (N-Nt), ammonium (N-Am) or urea (N-Ur) as the nitrogen (N) source at the same final N concentration (0.88 mM). Exponential growth phase cultures of the two diatoms were exposed to four different light regimes for 2 days: (UVAR) PAR (60 micromol quanta m-2 s-1) plus 8.22 W m-2 (unweighted) UVAR; (high UVBR) PAR (60 micromol quanta m-2 s-1) plus 1.04 W m-2 (unweighted) UVBR plus 13.73 W m-2 (unweighted) UVAR; (low UVBR) PAR (60 micromol quanta m-2 s-1) plus 0.19 W m-2 (unweighted) UVBR plus 2.76 W m-2 (unweighted) UVAR and (PAR) PAR (60 micromol quanta m-2 s-1) alone (control). No significant effects of N source on the growth rates of the two diatoms were detected. The maximum effective quantum yield of PSII, PhiPSIIe-max, and the initial slope of the light curve, alpha, of P. tricornutum and C. muelleri were all inhibited, whereas Ik was somewhat increased, as a consequence of 2 days of exposure to all the UVR treatments. Multiple factor ANOVA revealed that all the major fatty acids, both in P. tricornutum and C. muelleri, were influenced more strongly by N source than by UVR. The composition of saturated fatty acids (SFA), monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA) in P. tricornutum and C. muelleri exhibited almost the same pattern of variation with N source and UVR. The maximum value of SFA was found in the N-Am treatment, that of MUFA in the N-Nt treatment and for PUFA in the N-Ur treatment irrespective of the UV radiation. On the other hand, the impact of UVR resulted in an increase of PUFA and a reduction of SFA both in P. tricornutum and C. muelleri under all N sources.

Analysis of Variance↗

Mapping of nutrient-induced biochemical changes in living algal cells using synchrotron infrared microspectroscopy.

High quality Fourier transform infrared (FTIR) spectra were acquired from living Micrasterias hardyi cells maintained in an IR transparent flow-through cell using a FTIR microscope coupled to a synchrotron light source. Spectral maps of living, nutrient-replete cells showed band intensities consistent with the known location of the nucleus and the chloroplasts. These were very similar to maps acquired from fixed, air-dried cells. Bands due to lipids were lowest in absorbance in the region of the nucleus and highest in the chloroplast region and this trend was reversed for the absorbance of bands attributed to protein. Spectra acquired in 10 microm steps across living phosphorus-starved (P-starved) cells, repeated approximately every 30 min, were consistent over time, and bands correlated well with the known position of the nucleus and the observed chloroplasts, corroborating the observations with replete cells. Experiments in which missing nutrients were re-supplied to starved cells showed that cells could be maintained in a functional state in the flow-through cell for up to one day. Nitrogen-starved cells re-supplied with N showed an increase in lipid in all positions measured across the cell over a 23 h period of re-supply, with the largest increases occurring in positions where the chloroplasts were observed. Re-supply of phosphorus to P-starved cells produced no changes in bands attributable to lipid or protein. Due to their thin cell body ( approximately 12 microm) and large diameter ( approximately 300 microm) Micrasterias sp. make an ideal spectroscopic model to study nutrient kinetics in algal cells.

Eukaryota↗

A portable Raman acoustic levitation spectroscopic system for the identification and environmental monitoring of algal cells.

We report the coupling of a portable Raman spectrometer to an acoustic levitation device to enable environmental monitoring and the potential taxonomic identification of microalgae. Spectra of living cells were recorded at 785 nm using a fiber-optic probe coupled to a portable Raman spectrometer. The spectra exhibit an excellent signal-to-noise ratio and clearly show bands from chlorophyll a and beta-carotene. Spectra of levitated photobleached microalgae clearly show a reduction in chlorophyll a concentration relative to beta-carotene after 10 min of exposure to a quartz halogen lamp. Spectra recorded from levitated nitrogen-limited cells also show a significant reduction in bands associated with chlorophyll a, as compared to nitrogen-replete cells. To investigate the diagnostic capability of the technique, four species of microalgae were analyzed. Good quality spectra of all four species were obtained showing varying ratios of beta-carotene to chlorophyll. The combination of an acoustic levitation device and a portable Raman spectrometer shows potential as a taxonomic and environmental monitoring tool with direct application to field studies in remote environments.

Chlorophyta↗

Ultraviolet radiation has no effect on respiratory oxygen consumption or enhanced post-illumination respiration in three species of microalgae.

A 30-min exposure to UV-B radiation (1.1 Wm(-2), unweighted) from a xenon arc lamp caused pronounced inhibition (33-78%) of net photosynthetic oxygen production in three species of microalgae, Phaeodactylum tricornutum Bohlin, Dunaliella tertiolecta Butcher and Wolozynskia sp., however, no statistical differences (t-test, alpha=0.05) in dark-respiration rates were found between the control group and the UV-treated group, for any of the species tested. These results indicate: (i) that the respiratory processes responsible for oxygen consumption do not sustain any appreciable impairment registered in the first half-hour after ultraviolet radiation (UVR) exposure; and (ii) any change in respiration that may occur in response to increased repair demands is not detected in this period. Dark-respiration rates were observed to be significantly higher in all species tested (17-29%; t-test, alpha=0.05) following illumination with photosynthetically active radiation, compared to dark-respiration before illumination. This increase, interpreted as enhanced post-illumination respiration (EPIR), was observed in all three species. The magnitude of this increase was not affected by prior exposure to UVR.

Chlorophyta↗