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81.
Optical tweezers (infrared laser-based optical traps) have emerged as a powerful tool in molecular and cell biology. However, their usefulness has been limited, particularly in vivo, by the potential for damage to specimens resulting from the trapping laser. Relatively little is known about the origin of this phenomenon. Here we employed a wavelength-tunable optical trap in which the microscope objective transmission was fully characterized throughout the near infrared, in conjunction with a sensitive, rotating bacterial cell assay. Single cells of Escherichia coli were tethered to a glass coverslip by means of a single flagellum: such cells rotate at rates proportional to their transmembrane proton potential (. J. Mol. Biol. 138:541-561). Monitoring the rotation rates of cells subjected to laser illumination permits a rapid and quantitative measure of their metabolic state. Employing this assay, we characterized photodamage throughout the near-infrared region favored for optical trapping (790-1064 nm). The action spectrum for photodamage exhibits minima at 830 and 970 nm, and maxima at 870 and 930 nm. Damage was reduced to background levels under anaerobic conditions, implicating oxygen in the photodamage pathway. The intensity dependence for photodamage was linear, supporting a single-photon process. These findings may help guide the selection of lasers and experimental protocols best suited for optical trapping work. 相似文献
82.
83.
The tryptophan residues of bovine rhodopsin have been employed as intrinsic fluorescent probes for investigating the dynamics
of rhodopsin and of the thermal intermediates of its bleaching at low temperature. A fast depolarization of the fluorescence
of the tryptophans on the nanosecond time scale has been observed for all cases. The fluorescence spectra for the various
intermediates shift progressively to the red as a function of time following excitation with nanosecond light pulses. It is
inferred that rhodopsin possesses flexibility in the vicinity of its tryptophans that permits rapid structural fluctuations
to take place. 相似文献
84.
Metal concentrations in the gill, muscle and liver tissues of Labeo rosae from two impoundments, Loskop and Flag Boshielo dams on the Olifants River, were evaluated in 2011 to detect patterns in metal associations between tissues and impoundments. Elevated concentrations of Ba, Zn, B, Al, Si and Fe, relative to a pristine site in the catchment, were found in the muscle, liver and gill tissues at both impoundments. Molybdenum concentrations were exceptionally high in all tissues at Loskop Dam and in liver at Flag Boshielo Dam. No definite pattern in the ratio metal concentrations within, or between, fish tissues was identified. The expected trend, liver > gills > muscle, was found at both impoundments, but was less prominent at Loskop Dam. Metal concentrations in muscle of Loskop Dam fish were significantly higher than in those at Flag Boshielo Dam. The inverse was true for liver. The long-term impact of elevated metal concentrations on fish health at both impoundments raises concern. 相似文献
85.
Developing taste buds in the anterior mandibular floor of perihatching
chicks were studied by high voltage electron microscopic autoradiography in
order to identify proliferating gemmal cell types. Montaged profiles of 29
taste buds in five cases euthanized between embryonic day 21 and
posthatching day 2 were analyzed after a single [3H]thymidine injection
administered on embryonic day 16, 17 or 18. Results showed that dark cells
comprised 55% of identified (n = 900 cells) and 62% of labeled (n = 568
cells) gemmal cells as compared with light, intermediate, basal or
perigemmal bud cells. Dark cells had both a greater (P < 0.05) number of
labeled cells and a greater amount of label (grains/nucleus) than the other
four bud cell types, irrespective of injection day. The nuclear area
(micron 2) of dark cells was not significantly larger (P > 0.05) than
that of the other gemmal cell types and therefore cannot account for the
greater amount for label in the dark cells. Interestingly, only dark cells
showed a positive correlation (P < 0.003) between amount of label and
nuclear area. Results suggest that, during the perihatching period of
robust cell proliferation, dividing dark cells may give rise primarily, but
not exclusively, to dark cell progeny.
相似文献
86.
The major rivers of the South African ‘Lowveld’ (low-latitude savanna) suffer numerous impacts from upstream economic activities. Whereas monitoring these rivers is required to detect biodiversity losses, record pollution events and devise mitigation strategies, current monitoring programmes are inadequate. In 2009, the South African Earth Observation Network initiated an intensive long-term research programme on the Lowveld reaches of the Olifants River. Physico-chemical parameters, aquatic macroinvertebrates and fish abundances were recorded at four Lowveld sites in the Olifants River. We review six years of this programme. The results suggest deterioration in the ecological condition of the Olifants River with no discernible improvement through protected areas. Trends could not be detected. The parameters measured, sampling methods and/or sampling frequency might be responsible for the limited trends observed, or alternatively the results simply reflect stable conditions despite on-going pollution. Real time monitoring and an expansion in the parameters monitored would add value to the monitoring programme. 相似文献
87.
Galactose was specifically inserted into the carbohydrate moiety of rhodopsin by incubating retinal disk membranes with UDP-galactose: N-acetylglucosamine galactosyltransferase. The stoichiometry of labeling ranged from 1.2 to 1.8 (average = 1.5) residues of galactose per molecule of rhodopsin, indicating that some or all of the oligosaccharide chains of membrane-bound rhodopsin are readily accessible to enzymatic modification. These modified membranes were treated with galactose oxidase to generate an aldehyde at the C-6 position of the inserted galactose units. The enzymatically-oxidized membranes were then reacted with dansyl hydrazide to yield a fluorescent hydrazone which is sufficiently stable to permit spectroscopic analysis. This procedure for the specific attachment of a spectroscopic probe should be applicable to a wide variety of membrane glycoproteins. 相似文献