Effect of excitation wavelength on the Raman spectroscopy of the porcine photoreceptor layer from the area centralis.

Raman microscopy, based upon the inelastic scattering (Raman) of light by molecular species, has been applied as a specific structural probe in a wide range of biomedical samples. The purpose of the present investigation was to assess the potential of the technique for spectral characterization of the porcine outer retina derived from the area centralis, which contains the highest proportion of cone:rod cell ratio in the pig retina. METHODS: Retinal cross-sections, immersion-fixed in 4% (w/v) PFA and cryoprotected, were placed on salinized slides and air-dried prior to direct Raman microscopic analysis at three excitation wavelengths, 785 nm, 633 nm, and 514 nm. RESULTS: Raman spectra of each of the photoreceptor inner and outer segments (PIS, POS) and of the outer nuclear layer (ONL) of the retina acquired at 785 nm were dominated by vibrational features characteristic of proteins and lipids. There was a clear difference between the inner and outer domains in the spectroscopic regions, amide I and III, known to be sensitive to protein conformation. The spectra recorded with 633 nm excitation mirrored those observed at 785 nm excitation for the amide I region, but with an additional pattern of bands in the spectra of the PIS region, attributed to cytochrome c. The same features were even more enhanced in spectra recorded with 514 nm excitation. A significant nucleotide contribution was observed in the spectra recorded for the ONL at all three excitation wavelengths. A Raman map was constructed of the major spectral components found in the retinal outer segments, as predicted by principal component analysis of the data acquired using 633 nm excitation. Comparison of the Raman map with its histological counterpart revealed a strong correlation between the two images. CONCLUSIONS: It has been demonstrated that Raman spectroscopy offers a unique insight into the biochemical composition of the light-sensing cells of the retina following the application of standard histological protocols. The present study points to the considerable promise of Raman microscopy as a component-specific probe of retinal tissue.

Fe XIII emission lines in active region spectra obtained with the Solar Extreme-Ultraviolet Research Telescope and Spectrograph

Recent fully relativistic calculations of radiative rates and electron impact excitation cross-sections for FeXIII are used to generate emission-line ratios involving 3s23p2-3s3p3 and 3s23p2-3s23p3d transitions in the 170-225 and 235-450 Å wavelength ranges covered by the Solar Extreme-Ultraviolet Research Telescope and Spectrograph (SERTS). A comparison of these line ratios with SERTS active region observations from rocket flights in 1989 and 1995 reveals generally very good agreement between theory and experiment. Several new FeXIII emission features are identified, at wavelengths of 203.79, 259.94, 288.56 and 290.81 Å. However, major discrepancies between theory and observation remain for several FeXIII transitions, as previously found by Landi and others, which cannot be explained by blending. Errors in the adopted atomic data appear to be the most likely explanation, in particular for transitions which have 3s23p3d1D2 as their upper level. The most useful FeXIII electron-density diagnostics in the SERTS spectral regions are assessed, in terms of the line pairs involved being (i) apparently free of atomic physics problems and blends, (ii) close in wavelength to reduce the effects of possible errors in the instrumental intensity calibration, and (iii) very sensitive to changes in Ne over the range 108-1011cm-3. It is concluded that the ratios which best satisfy these conditions are 200.03/202.04 and 203.17/202.04 for the 170-225 Å wavelength region, and 348.18/320.80, 348.18/368.16, 359.64/348.18 and 359.83/368.16 for 235-450 Å.

Macroturbulent and rotational broadening in the spectra of B-type supergiants.

The absorption-line spectra of early B-type supergiants show significant broadening that implies that an additional broadening mechanism (characterized here as `macroturbulence') is present in addition to rotational broadening. Using high-resolution spectra with signal-to-noise ratios of typically 500, we have attempted to quantify the relative contributions of rotation and macroturbulence, but even with data of this quality significant problems were encountered. However, for all our targets, a model where macroturbulence dominates and rotation is negligible is acceptable; the reverse scenario leads to poor agreement between theory and observation. Additionally, there is marginal evidence for the degree of broadening increasing with line strength, possibly a result of the stronger lines being formed higher in the atmosphere. Acceptable values of the projected rotational velocity are normally less than or equal to 50 km s-1, which may also be a typical upper limit for the rotational velocity. Our best estimates for the projected rotational velocity are typically 10-20 km s-1 and hence compatible with this limit. These values are compared with those predicted by single star evolutionary models, which are initially rapidly rotating. It is concluded that either these models underestimate the rate of rotational breaking or some of the targets may be evolving through a blue loop or are binaries.

A non-LTE analysis of the spectra of two narrow lined main sequence stars in the SMC

An analysis of high-resolution VLT/UVES spectra of two B-type main sequence stars, NGC 346-11 and AV 304, in the Small Magellanic Cloud (SMC), has been undertaken, using the non-LTE tlusty model atmospheres to derive the stellar parameters and chemical compositions of each star. The chemical compositions of the two stars are in reasonable agreement. Moreover, our stellar analysis agrees well with earlier analyses of H II regions. The results derived here should be representative of the current base-line chemical composition of the SMC interstellar medium as derived from B-type stars.

Extreme ultraviolet transitions of Fe XXI in solar, stellar and laboratory spectra

Recent R-matrix calculations of electron impact excitation rates for transitions among the 2s(2)2p(2), 2s2p(3) and 2p(4) levels of Fe XXI are used to derive theoretical electron density (N-e) sensitive emission-line ratios involving 2S2(2)p(2)-2s2p(3) transitions in the similar to 98-146 Angstrom wavelength range. A comparison of these with observations from the PLT tokamak plasma, for which the electron density has been independently determined, reveals generally very good agreement between theory and experiment, and in some instances removes discrepancies found previously. The observed Fe XXI ratios for a solar flare, obtained with the OSO-5 satellite, imply electron densities which are consistent, with discrepancies that do not exceed 0.2 dex. In addition, the derived values of N-e are similar to those estimated for the high-temperature regions of other solar flares. The good agreement between theory and observation, in particular for the tokamak spectra, provides experimental support for the accuracy of the present line-ratio calculations, and hence for the atomic data on which they are based.

Emission lines of [Cl II] in the optical spectra of gaseous nebulae

Recent R-matrix calculations of electron impact excitation rates among the 3s(2)3p(4) levels of Cl II are used to derive the nebular emission-line intensity ratios R-1=I(6161.8 Angstrom)/I(8578.7 Angstrom) and R-2=I(6161.8 Angstrom)/I(9123.6 Angstrom) as a function of electron temperature (T-e) and density (N-e). The ratios are found to be very sensitive to changes in T-e but not N-e for densities lower than 10(5) cm(-3). Hence, they should, in principle, provide excellent optical T-e diagnostics for planetary nebulae. The observed values of R-1 and R-2 for the planetary nebulae NGC 6741 and IC 5117, measured from spectra obtained with the Hamilton echelle spectrograph on the 3 m Shane Telescope, imply temperatures in excellent agreement with those derived from other diagnostic lines formed in the same region of the nebula as [Cl II]. This provides some observational support for the accuracy of the [Cl II] line ratio calculations and hence the atomic data on which they are based. The [Cl II] 8578.7 and 9123.6 Angstrom lines are identified for the first time (to our knowledge) in a high-resolution spectrum of the symbiotic star RR Telescopii, obtained with the University College London Echelle Spectrograph on the 3.9 m Anglo- Australian Telescope. However, the 6161.8 Angstrom feature is unfortunately too weak to be identified in the RR Telescopii observations, consistent with its predicted line strength.

SiIX emission lines in spectra obtained with the Solar EUV Research Telescope and Spectrograph (SERTS)

Theoretical electron-density-sensitive emission line ratios involving 2s(2)2p(2)-2s2p(3) transitions in Si IX between 223 and 350 Angstrom are presented. A comparison of these with an extensive dataset of solar-active-region, quiet-Sun, subflare and off-limb observations, obtained during rocket flights by the Solar EUV Research Telescope and Spectrograph (SERTS), reveals generally very good agreement between theory and experiment. This provides support for the accuracy of the line- ratio diagnostics, and hence the atomic data on which they are based. In particular, the density-sensitive intensity ratio I (258.10 Angstrom)/ I (349.87 Angstrom) offers an especially promising diagnostic for studies of coronal plasmas, as it involves two reasonably strong emission lines and varies by more than an order of magnitude over the useful density range of 10(9)-10(11) cm(-3). The 2s(2)2p(2) S-1(0) - 2s2p(3) P-1(1) transition at 259.77 Angstrom is very marginally identified for the first time in the SERTS database, although it has previously been detected in solar flare observations.

Emission lines of Na-like ions in spectra obtained with the Solar EUV Research Telescope and Spectrograph (SERTS)

Theoretical emission-line ratios involving transitions in the 236-412 Angstrom wavelength range are presented for the Na-like ions Ar viii, Cr xiv, Mn xv, Fe xvi, Co xvii, Ni xviii and Zn xx. A comparison of these with an extensive data set of the solar active region, quiet-Sun, subflare and off-limb observations, obtained during rocket flights by the Solar EUV Research Telescope and Spectrograph (SERTS), reveals generally very good agreement between theory and experiment. This indicates that most of the Na-like ion lines are reliably detected in the SERTS observations, and hence may be employed with confidence in solar spectral analyses. However, the features in the SERTS spectra at 236.34 and 300.25 Angstrom, originally identified as the Ni xviii 3p (2) P-3/2 -3d (2) D- 3/2 and Cr xiv 3p (2) P-3/2 -3d (2) D-5/2 transitions, respectively, are found to be due to emission lines of Ar xiii (236.34 Angstrom) and possibly S v or Ni vi (300.25 Angstrom). The Co xvii 3s (2) S-3p (2) P-3/2 line at 312.55 Angstrom is always badly blended with an Fe xv feature at the same wavelength, but Mn xv 3s (2) S-3p (2) P-1/2 at 384.75 Angstrom may not always be as affected by second-order emission from Fe xii 192.37 Angstrom as previously thought. On the other hand, we find that the Zn xx 3s (2) S-3p (2) P-3/2 transition can sometimes make a significant contribution to the Zn xx/Fe xiii 256.43- Angstrom blend, and hence care must be taken when using this feature as an Fe xiii electron density diagnostic. A line in the SERTS-89 active region spectrum at 265.00 Angstrom has been re-assessed, and we confirm its identification as the Fe xvi 3p (2) P-3/2 -3d (2) D-3/2 transition.