共查询到20条相似文献,搜索用时 31 毫秒
1.
We have previously developed a model which includes energy and phasechanging collisional relaxations in a system of chemically reacting molecules absorbing light from a monochromatic field while immersed in an inert thermal bath. Some aspects of the model are presented here which relate to scattered light measurements on macromolecular systems. We predict that in the presence of low-intensity laser light the elastic component of the scattered light will be broadened by the rate constant for the chemical reaction. In the presence of high-intensity laser light, the scattered field may contain, in addition to the Rayleigh scattered light, two sidebands symmetrically displaced to either side of the Rayleigh band; the magnitude of the displacement is a function of the laser intensity. The Rayleigh band width is a direct measure of the phase relaxation time, and the sideband widths are a measure of the energy and phase relaxation times. We discuss several experimental systems in which sideband scattering data might be used to provide information related to relaxation and reaction mechanisms in macromolecular systems of biological interest; bound enzyme-substrate and enzyme-dye systems can be investigated as can vibrational energy transfer on membrane-bound systems. Some numerical computations are included for the magnitude of the sideband displacement and scattered intensities. 相似文献
2.
A system for imaging transverse distribution of scattered light and chlorophyll fluorescence in intact rice leaves 总被引:5,自引:1,他引:4
K. TAKAHASHI K. MINEUCHI T. NAKAMURA M. KOIZUMI H. KANO 《Plant, cell & environment》1994,17(1):105-110
In order to examine the transverse distribution of scattered light and chlorophyll fluorescence in intact rice leaves, a micro-fluorescence imaging system was devised using a microscope, a CCD camera with an image intensifier, an Ar and a He-Ne laser light source, an image processor, and a microcomputer. A laser light was projected vertically on to the surface of a rice leaf segment at a cut-edge, and scattered light and induced fluorescence were observed at the cut-section from a 90° angle to the axis of the laser beam. The intensity of scattered light showed a maximum at several micrometres depth from the leaf surface and a steep gradient afterwards. Fluorescence reached a maximum crossing with the decline curve of the scattered light. The maximum of fluorescence measured at 741 nm was observed at a greater depth from the leaf surface than that at 687 nm, suggesting that part of the fluorescence of the longer wavelength was emitted due to absorption of fluorescence of the shorter wavelength. Profiles of the scattered light and the chlorophyll fluorescence depended on leaf anatomy. 相似文献
3.
A method is described for estimating the fractional contribution of light scattered from the excitation lamp to the normalized raw fluorescence lifetime curve. The method depends on the ratio of the slope of the normalized light scatter spectrum to the slope of the normalized raw fluorescence spectrum in the vicinity of the intersection of the two spectra. The correction for scattered light is made prior to deconvolution, and hence, has the advantage of being independent of the method selected to calculate the true fluorescence life-time spectrum. It is simple and does not require a computer. Tested against curves synthesized from known additions of scattered light to fluorescence spectra exhibiting mono-, bi-, or triexponential decay, it yielded small absolute errors. 相似文献
4.
As exciting light in a scanning confocal microscope encounters a cell and its subcellular components, it is refracted and scattered. A question arises as to what proportion of the exciting light is scattered by subcellular structures and whether cells in the vicinity of the imaged area, i.e., cells that are not directly illuminated by the laser beam, can be affected by either an exposure to scattered light and ensuing phototoxic reactions, or by the products of photoactivated reactions diffusing out of the directly illuminated area. We have designed a technique, which allows us to detect subtle cell photodamage and estimate the extent and range of phototoxic effects inflicted by interaction between scattered exciting light and fluorescent probes in the vicinity of the illuminated area. The technique is based on detecting an increased influx of acridine orange into photodamaged cells, which is manifested by a change of color. We demonstrate that phototoxic effects can be exerted not only on the illuminated cell, but also on fluorescently labeled neighboring cells. The damage inflicted on neighbors is due to exposure to light scattered by the imaged (i.e., directly illuminated) cell, but not phototoxic products diffusing out of the directly illuminated area. When light encounters a cell nucleus, scattering is so intense that photodamage can be inflicted even on fluorescently labeled cells located within a radius of approximately 90 microm, i.e., several cell diameters away. This range of scattering is comparable with that caused by the glass bead resting on a coverslip (up to 120 microm). The intense scattering of exciting light imposes limits on FRAP, FLIP, and other techniques employing high intensity laser beams. 相似文献
5.
The intensity of light scattered at 90° to the incident beam and the effective hydrodynamic radii of mitochondria incubated under a variety of conditions have been measured. Addition of high concentrations of uncouplers to respiring mitochondria resulted in a decrease in scatter which was not due to swelling. Addition of valinomycin to mitochondria depleted of substrate in K+-free medium produced an increase in scatter that was not due to shrinking. It is concluded that changes in the intensity of scattered light are not reliable indices of changes of volume of mitochondria, and that changes in conformation with changes in metabolic state dominate changes in light scatter. A molecular mechanism for the effect of metabolic state upon the scattered intensity is suggested. 相似文献
6.
Light gradients and spectral regime were measured in Lactuca sativa L. cv. Grand Rapids achenes using fiber optic microsensors. The distribution of scattered light across lettuce achenes was linear for 660 and 730 nm and non-linear for 450 nm light. Spectra for scattered light within intact achenes also showed a non-linear increase with wavelength. The preferential attenuation of blue light by the pericarp and seed explains in part the relative ineffectiveness of blue light with respect to red in triggering germination of lettuce. Calculated action spectra for phytochrome-stimulated germination agree closely in the red with experimentally derived action spectra; however, there is little agreement within the blue. 相似文献
7.
Discrimination of eukaryotic phytoplankton cell types from light scatter and autofluorescence properties measured by flow cytometry 总被引:5,自引:0,他引:5
Flow cytometric methods for recognizing several groups of eukaryotic marine phytoplankton were tested using 26 laboratory cultures. Each culture was divided into three aliquots, and these samples were analyzed for 1) Coulter volume; 2) light scatter (magnitude and polarization properties of forward scattered light and magnitude of right-angle scattered light) and autofluorescence emission (phycoerythrin and chlorophyll); and 3) autofluorescence excitation (by 488 nm and 515 nm light). Three kinds of cells could be easily distinguished from others in the culture collection: 1) The two cryptophytes and the rhodophyte had high phycoerythrin/chlorophyll ratios; 2) the two coccolithophores depolarized forward scattered light; and 3) the two pennate diatoms scattered only a relatively small amount of light in the forward direction compared with that at right angles. Mean chlorophyll fluorescence excited by blue light relative to that excited by green light was highest in the four chlorophytes, but there was overlap between some of these and some other kinds of cells. Unresolved cell types included centric diatoms, dinoflagellates, and naked coccolithophores. Forward light scatter and Coulter volume were closely related (except for the pennate diatoms) over a range of about 0.01 to 30 pL (equivalent spherical diameter about 3 to 40 microns), according to a logarithmic function. 相似文献
8.
We have obtained a rotational diffusion coefficient of the 70S ribosome isolated from Escherichia-coli (MRE-600), from the depolarized light scattering spectrum measured by photon correlation spectroscopy. The intensity correlation function of depolarized scattered light contains contributions due to multiple scattered and anisotropy scattered light from the ribosomal particle. We discuss extensively the subtraction procedure used to obtain the rotational correlation time from the experimental correlation function. We have also obtained the translational diffusion coefficient from the same sample by determining the polarized correlation function. The hydrodynamic radius determined from the rotational diffusion coefficient is only slightly larger than the radius obtained from the translational diffusion coefficient. Therefore the ribosomal particle has a non-spherical shape. This conclusion, however, could be impaired by the effect of free draining of the ribosome. 相似文献
9.
We have obtained a rotational diffusion coefficient of the 70S ribosome isolated from Escherichia-coli (MRE-600), from the depolarized light scattering spectrum measured by photon correlation spectroscopy. The intensity correlation function of depolarized scattered light contains contributions due to multiple scattered and anisotropy scattered light from the ribosomal particle. We discuss extensively the subtraction procedure used to obtain the rotational correlation from the time from the experimental correlation function. We have also obtained the translational diffusion coefficient from the same sample by determining the polarized correlation function. The hydrodynamic radius determined from the rotational diffusion coefficient is only slightly larger than the radius obtained from the translational diffusion coefficient. Therefore the ribosomal particle has a non-spherical shape. This conclusion, however, could be impaired by the effect of free draining of the ribosome. 相似文献
10.
John W. Preiss 《Biophysical journal》1968,8(2):199-209
Pancreatic ribonuclease was irradiated in the dried state with electrons and then added to acetate buffer solutions that contained different concentrations of polyribonucleotides. Qualitatively similar results were obtained by adding a combination of unirradiated ribonuclease and lysozyme to such solutions. Such solutions scatter light strongly, and the intensity of the scattered light changes with time after mixing. The angular distribution of the scattered light was obtained as a function of time and compared with the rates at which hydrolysis products were formed. The turbidity of the solutions increases rapidly with time at the lower polyribonucleotide concentrations, and seems to result from a complex between inactive ribonuclease, or lysozyme, and oligonucleotides that appear during enzymic hydrolysis of the polynucleotides. The dissymmetry of the scattered light is approximately 5, indicating that the scattering centers are, if spherical, about 1500 A in diameter. The turbidities are remarkably high when one considers the low concentrations of protein and nucleic acid materials that are used. 相似文献
11.
Light and chlorophyll gradients within Cucurbita cotyledons 总被引:1,自引:4,他引:1
Abstract. Measurement of light within 10–14-d-old green and etiolated Cucurbita pepo cotyledons were made with fibre-optic microprobes to assess the influence of chlorophyll distribution and anatomical variations in mesophyll cell type (spongy versus palisade) on internal light pattern. More than 50% of the pigment in green cotyledons occurred in the upper (adaxial) 300 μm and this gradient strongly influenced the internal propagation of 680 nm light. When the upper (adaxial) surface was irradiated with 680 nm light, almost complete absorption occurred within the first 400 μm (palisade) of approximately 1200-μm-thick cotyledons. In contrast, when lower (abaxial) surfaces were irradiated with 680 nm light, penetration extended throughout the spongy mesophyll to about the 700 μm depth. Measurements of collimaled and scattered light gradients at 550, 680 and 750 nm indicated that collimaled light was rapidly scattered by mesophyll cells. In cotyledons irradiated on the upper surface, spongy mesophyll cells received only scattered light. Furthermore, comparisons of scattered light gradients obtained from cotyledons irradiated on upper and lower surfaces suggested that spongy mesophyll cells scatter light more effectively than palisade cells, probably due to the greater proportion of intercellular air spaces in spongy mesophyll tissue. These data also indicate that both the spectral quality and quantity of light incident on palisade versus spongy mesophyll cells differs, perhaps contributing to developmental and physiological differences between these two mesophyll cell types. 相似文献
12.
T. Matsuura S. Gorti T. Tanaka Yoshiaki Hara Mototsugu Saishin 《European biophysics journal : EBJ》1999,28(5):357-368
From observations of the dynamics of light scattered by the cornea, intensity autocorrelation func-tions that revealed two
independent diffusion coefficients, D (fast) = 2.4±0.2×10–7 cm2/s and D (slow) = 9.4±1.3× 10–9 cm2/s, were obtained. The diffusion coefficients were found to be statistically independent of the position and depth on the
lateral surface of the cornea from which the scattered light was sampled. The slow diffusion coefficients obtained from light
sampled from within cross-sections of the cornea were, however, measurably different. Diffusion coefficients obtained independently
from observations of the kinetics of corneal swelling for comparison were found to be several orders of magnitude greater
than those obtained from light scattering. The large disparity in the diffusion coefficients obtained from the two independent
methods invoked the possibility that the lamellar layers within the cornea behave as individual gel sheets. Irrespective of
this additional hypothesis, divergent behavior in the measured total scattered light intensities and diffusion coefficients
upon varying external conditions, such as temperature or pressure (stretching), was observed. Namely, a slowing down of the
dynamic modes accompanied by increased “static” scattered light intensities was observed. Although the slowing down of the
dynamic modes is possibly indicative of the reduced affinity of protein binding to the gel matrix that “softens” the gel,
the divergent behavior in the scattered light intensities and diffusion coefficients is, however, more characteristic of a
phase transition. In addition, the divergent behavior in the scattered light intensities and diffusion coefficients was reversible
up to a critical temperature (∼55 °C) or stretching (∼16%).
Received: 18 March 1998 / Revised version: 4 February 1999 / Accepted: 4 February 1999 相似文献
13.
Identification of the scattering elements responsible for lens opacification in cold cataracts. 总被引:6,自引:2,他引:4
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Using both quasi-elastic light scattering spectroscopy and angular dissymmetry in the intensity of the scattered light, we examined the onset of turbidity for intact calf lenses and for isolated nuclear cytoplasm. In the case of the nuclear cytoplasm these measurements demonstrate the presence of two kinds of scatterers: small units of approximately 100-A radius and larger elements whose size is distributed around 1,500 A. As the temperature is decreased towards the cold cataract temperature, the intensity of light scattered by the small units stays almost constant while the intensity scattered by the large elements increase very strongly. The opacification of the lens cytoplasm produced by decreasing the temperature results principally from an increase in the concentration of the large scattering elements. For the intact nucleus the situation is qualitatively similar, but the mean size of the large scattering elements shows a more substantial increase than in the isolated cytoplasm as temperature is lowered towards the cold cataract temperature. 相似文献
14.
A Porath-Furedi 《Cytometry》1983,4(3):263-267
The measurement of the fluctuations of the scattered light were the source of information on the flatness of erythrocytes. Data on cell volume, together with the measure of scattered light fluctuations, defined the cell shape. The measurements were repeated in a wide range of osmotic pressures and pH values in order to affect both the hemoglobin and cytoskeleton. Major volume changes were detected at low osmotic pressures and pH. The erythrocyte volume was determined by electronic volume measurement. The cell flatness is maximal at physiological conditions. 相似文献
15.
16.
Calculations based on the known dimensions of bull spermatozoa show that the scattered light intensity is strongly dependent upon the relative orientation of the particle to the incident beam. The magnitude of this effect of apparently much greater than for other systems where motility has been investigated by dynamic light scattering. The calculations show that the scattering source can be approximated by a small spinning mirror, and consequently the greatest light intensity at the detector results from cells swimming in a direction perpendicular to the scattering vector. The calculations are in substantial agreement with photographic observations, as well as direct measurements of the scattered intensity. Previous treatments of dynamic light scattering from swimming bull spermatozoa based on point scattering models are shown to be incorrect. 相似文献
17.
Depolarization of circularly polarized light scattered from biological tissues depends on structural changes in cell nuclei, which can provide valuable information for differentiating cancer tissues concealed in healthy tissues. In this study, we experimentally verified the possibility of cancer identification using scattering of circularly polarized light. We investigated the polarization of light scattered from a sliced biological tissue with various optical configurations. A significant difference between circular polarizations of light scattered from cancerous and healthy tissues is observed, which is sufficient to distinguish a cancerous region. The line-scanning experiments along a region incorporating healthy and cancerous parts indicate step-like behaviors in the degree of circular polarization corresponding to the state of tissues, whether cancerous or normal. An oblique and perpendicular incidence induces different resolutions for identifying cancerous tissues, which indicates that the optical arrangement can be selected according to the priority of resolution. 相似文献
18.
This study was concerned with the quantitative evaluation of dark field light scattering by sedimented erythrocytes of banked human blood samples. Due to considerable variability of both appearance and amount of scattered light the discocyte group had to be subdivided into discocyte I and discocyte II. The mean intensity of scattered light increased about three fold from discocyte II to echinocytes I, II, III, sphaeroechinocyte, and sphaerocyte. On the other hand the average light scattering intensity of discocytes I exceeded that of discocytes II about 2.5 times, with individual data varying over a wide range. There was a rapid disappearing of discocytes I correlated with time of storage. Therefore it is concluded that discocytes I represent the initial stage of erythrocytes transforming under banking conditions. 相似文献
19.
The effects of scattered actinic radiation on photomultipliers (Hamamatsu R-562) were investigated. Using cotton-wool to model dense biological preparations, it was found that the scattered actinic radiation received by the photomultiplier gives rise to phytochrome-like signals. This demonstrated the necessity to shield the photomultiplier from scattered actinic light for sensitive measurements with light-scattering preparations. 相似文献
20.
We describe an instrument for monitoring either transmitted or scattered light intensity, or both, simultaneously on up to eight channels. The use of a laser light source (at 632.8-nm wavelength) provides high accuracy and dynamic range: optical density can be measured from 0.0004 up to 6, and a scattered light fraction down to 10(-6) can be resolved. Built-in thermostat and magnetic stirrers allow precise monitoring of aqueous microbial growth over a practical range of 4 orders of magnitude of cell concentration. 相似文献