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The Journal of Cell Biology Nov 1969By applying microspectrophotometry to the sporangiophore of Phycomyces blakesleeanus wild-type and the albino car-10(-) type II, absorption spectra were obtained for 1-...
By applying microspectrophotometry to the sporangiophore of Phycomyces blakesleeanus wild-type and the albino car-10(-) type II, absorption spectra were obtained for 1- to 5-day cultures. Spectra in the growing-zone of the wild-type during Stage IVb, taken from 0.1 to 3 mm below the base of the sporangium, show two distinctly different spectra: one is more characteristic of a carotene, the other of a flavin. Combined, these absorption spectra reproduce closely the action spectrum. For the albino car-10(-), which is deficient in carotenes, only the spectrum characteristic of lumichrome or a reduced flavin was found. A c-type cytochrome was isolated from both strains which, if coupled with a flavin, could permit a photoreversible oxidation-reduction system. Birefringent crystals were observed to be aligned in the growing zone in which the photoreceptor is believed to lie. Micro-spectrophotometry of these crystals shows absorption peaks similar to those of riboflavin crystals.
Topics: Birefringence; Carotenoids; Cytochromes; Flavins; Fungi; Light; Microscopy, Electron; Pigmentation; Riboflavin; Sensory Receptor Cells; Spectrophotometry; Spores
PubMed: 5344152
DOI: 10.1083/jcb.43.2.354 -
The Journal of Experimental Biology May 2016Crocodilians are apex amphibious predators that occupy a range of tropical habitats. In this study, we examined whether their semi-aquatic lifestyle and ambush hunting...
Crocodilians are apex amphibious predators that occupy a range of tropical habitats. In this study, we examined whether their semi-aquatic lifestyle and ambush hunting mode are reflected in specific adaptations in the peripheral visual system. Design-based stereology and microspectrophotometry were used to assess spatial resolving power and spectral sensitivity of saltwater (Crocodylus porosus) and freshwater crocodiles (Crocodylus johnstoni). Both species possess a foveal streak that spans the naso-temporal axis and mediates high spatial acuity across the central visual field. The saltwater crocodile and freshwater crocodile have a peak spatial resolving power of 8.8 and 8.0 cycles deg(-1), respectively. Measurement of the outer segment dimensions and spectral absorbance revealed five distinct photoreceptor types consisting of three single cones, one twin cone and a rod. The three single cones (saltwater/freshwater crocodile) are violet (424/426 nm λmax), green (502/510 nm λmax) and red (546/554 nm λmax) sensitive, indicating the potential for trichromatic colour vision. The visual pigments of both members of the twin cones have the same λmax as the red-sensitive single cone and the rod has a λmax at 503/510 nm (saltwater/freshwater). The λmax values of all types of visual pigment occur at longer wavelengths in the freshwater crocodile compared with the saltwater crocodile. Given that there is a greater abundance of long wavelength light in freshwater compared with a saltwater environment, the photoreceptors would be more effective at detecting light in their respective habitats. This suggests that the visual systems of both species are adapted to the photic conditions of their respective ecological niche.
Topics: Alligators and Crocodiles; Animals; Cell Count; Cell Differentiation; Color Vision; Fresh Water; Microspectrophotometry; Photoreceptor Cells, Vertebrate; Retinal Ganglion Cells; Seawater; Vision, Ocular
PubMed: 27208035
DOI: 10.1242/jeb.135673 -
Current Biology : CB Feb 2015The mesopelagic habitat is a vast space that lacks physical landmarks and is structured by depth, light penetration, and horizontal currents. Solar illumination is...
The mesopelagic habitat is a vast space that lacks physical landmarks and is structured by depth, light penetration, and horizontal currents. Solar illumination is visible in the upper 1,000 m of the ocean, becoming dimmer and spectrally filtered with depth-generating a nearly monochromatic blue light field. The struggle to perceive dim downwelling light and bioluminescent sources and the need to remain unseen generate contrasting selective pressures on the eyes of mesopelagic inhabitants. Hyperiid amphipods are cosmopolitan members of the mesopelagic fauna with at least ten different eye configurations across the family-ranging from absent eyes in deep-living species to four enlarged eyes in mesopelagic individuals. The hyperiid amphipod Paraphronima gracilis has a pair of bi-lobed apposition compound eyes, each with a large upward-looking portion and a small lateral-looking portion. The most unusual feature of the P. gracilis eye is that its upward-looking portion is resolved into a discontinuous retina with 12 distinct groups, each serving one transverse row of continuously spaced facets. On the basis of eye morphology, we estimated spatial acuity (2.5° ± 0.11°, SEM; n = 25) and optical sensitivity (30 ± 3.4 μm(2) ⋅ sr, SEM; n = 25). Microspectrophotometry showed that spectral sensitivity of the eye peaked at 516 nm (±3.9 nm, SEM; n = 6), significantly offset from the peak of downwelling irradiance in the mesopelagic realm (480 nm). Modeling of spatial summation within the linear retinal groups showed that it boosts sensitivity with less cost to spatial acuity than more typical configurations.
Topics: Amphipoda; Animals; Compound Eye, Arthropod; Microscopy, Electron, Scanning; Visual Acuity; Visual Perception; Zooplankton
PubMed: 25601548
DOI: 10.1016/j.cub.2014.12.010 -
Vision Research Oct 2016The guppy (Poecilia reticulata) shows remarkable variation of photoreceptor cells in the retina, especially those sensitive to middle-to-long wavelengths of light....
The guppy (Poecilia reticulata) shows remarkable variation of photoreceptor cells in the retina, especially those sensitive to middle-to-long wavelengths of light. Microspectrophotometry (MSP) has revealed varying "green", "green-yellow" and "yellow" cone cells among guppies in Trinidad and Venezuela (Cumana). In the guppy genome, there are four "long-wave" opsin loci (LWS-1, -2, -3 and -4). Two LWS-1 alleles have potentially differing spectral sensitivity (LWS-1/180Ser and LWS-1/180Ala). In addition, two "middle-wave" loci (RH2-1 and -2), two "short-wave" loci (SWS2-A and -B), and a single "ultraviolet" locus (SWS1) as well as a single "rhodopsin" locus (RH1) are present. However, the absorption spectra of these photopigments have not been measured directly and the association of cell types with these opsins remains speculative. In the present study, we reconstituted these opsin photopigments in vitro. The wavelengths of maximal absorbance (λmax) were 571nm (LWS-1/180Ser), 562nm (LWS-1/180Ala), 519nm (LWS-3), 516nm (LWS-2), 516nm (RH2-1), 476nm (RH2-2), 438nm (SWS2-A), 408nm (SWS2-B), 353nm (SWS1) and 503nm (RH1). The λmax of LWS-3 is much shorter than the value expected (560nm) from the "five-sites" rule. The two LWS-1 alleles could explain difference of the reported MSP λmax values for the yellow cone class between Trinidad and Cumana guppies. Absence of the short-wave-shifted LWS-3 and the green-yellow cone in the green swordtail supports the hypothesis that this cell class of the guppy co-expresses the LWS-1 and LWS-3. These results reveal the basis of variability in the guppy visual system and provide insight into the behavior and ecology of these tropical fishes.
Topics: Alleles; Animals; Color Perception; Cone Opsins; Gene Expression Profiling; Poecilia; Retinal Cone Photoreceptor Cells; Retinal Pigments
PubMed: 27476645
DOI: 10.1016/j.visres.2016.06.013 -
AAPS PharmSciTech Jun 2013A new, simple, inexpensive, and rapid 96-well plate UV spectrophotometric method was developed and validated for the quantification of compound 48/80 (C48/80) associated...
A new, simple, inexpensive, and rapid 96-well plate UV spectrophotometric method was developed and validated for the quantification of compound 48/80 (C48/80) associated with particles. C48/80 was quantified at 570 nm after reaction with acetaldehyde and sodium nitroprusside in an alkaline solution (pH 9.6). The method was validated according to the recommendations of the ICH Guidelines for specificity, linearity, range, accuracy, precision, and detection and quantification limits (DL and QL). All the validation parameters were assessed in three different solvents, i.e., deionized water, blank matrix of chitosan nanoparticles, and blank matrix of chitosan/alginate nanoparticles. The method was found to be linear in the concentration range of 5 to 160 μg/ml (R(2)>0.9994). Intraday and interday precision was adequate, with relative standard deviation lower than those given by the Horwitz equation. The mean recoveries of C48/80 from spiked samples ranged between 98.1% and 105.9% for calibration curves done with the blank matrices and between 89.3% and 103.3% for calibration curves done with water, respectively. The DL were lower than 1.01 μg/ml and the QL were lower than 3.30 μg/ml. The results showed that the developed method is sensitive, linear, precise, and accurate for its intended use, with the additional advantages of being cost-effective and time-effective, allowing the use of small-volume samples, and the simultaneous analysis of a large number of samples. The proposed method was already successfully applied to evaluate the loading efficacy of C48/80 chitosan-based nanoparticles and can be easily applied during the development of other C48/80-based formulations.
Topics: Acetaldehyde; Adjuvants, Immunologic; Alginates; Calibration; Chemistry, Pharmaceutical; Chitosan; Drug Carriers; Equipment Design; Hydrogen-Ion Concentration; Limit of Detection; Linear Models; Microspectrophotometry; Miniaturization; Nanoparticles; Nitroprusside; Reference Standards; Reproducibility of Results; Solvents; Spectrophotometry, Ultraviolet; Technology, Pharmaceutical; Water; p-Methoxy-N-methylphenethylamine
PubMed: 23468340
DOI: 10.1208/s12249-013-9950-4 -
Experimental Dermatology Jan 2014Plant-derived oils consisting of triglycerides and small amounts of free fatty acids (FFAs) are commonly used in skincare regimens. FFAs are known to disrupt skin...
Plant-derived oils consisting of triglycerides and small amounts of free fatty acids (FFAs) are commonly used in skincare regimens. FFAs are known to disrupt skin barrier function. The objective of this study was to mechanistically study the effects of FFAs, triglycerides and their mixtures on skin barrier function. The effects of oleic acid (OA), glyceryl trioleate (GT) and OA/GT mixtures on skin barrier were assessed in vivo through measurement of transepidermal water loss (TEWL) and fluorescein dye penetration before and after a single application. OA's effects on stratum corneum (SC) lipid order in vivo were measured with infrared spectroscopy through application of perdeuterated OA (OA-d34 ). Studies of the interaction of OA and GT with skin lipids included imaging the distribution of OA-d34 and GT ex vivo with IR microspectroscopy and thermodynamic analysis of mixtures in aqueous monolayers. The oil mixtures increased both TEWL and fluorescein penetration 24 h after a single application in an OA dose-dependent manner, with the highest increase from treatment with pure OA. OA-d34 penetrated into skin and disordered SC lipids. Furthermore, the ex vivo IR imaging studies showed that OA-d34 permeated to the dermal/epidermal junction while GT remained in the SC. The monolayer experiments showed preferential interspecies interactions between OA and SC lipids, while the mixing between GT and SC lipids was not thermodynamically preferred. The FFA component of plant oils may disrupt skin barrier function. The affinity between plant oil components and SC lipids likely determines the extent of their penetration and clinically measurable effects on skin barrier functions.
Topics: Adult; Body Water; Dermatologic Agents; Epidermis; Female; Humans; In Vitro Techniques; Lipid Metabolism; Microspectrophotometry; Oleic Acid; Plant Oils; Skin Absorption; Triolein; Young Adult
PubMed: 24372651
DOI: 10.1111/exd.12296 -
Journal of Biomedical Optics Apr 2012A tutorial article is presented for the use of linear and nonlinear Raman microspectroscopies in biomedical diagnostics. Coherent anti-Stokes Raman scattering (CARS) is...
A tutorial article is presented for the use of linear and nonlinear Raman microspectroscopies in biomedical diagnostics. Coherent anti-Stokes Raman scattering (CARS) is the most frequently applied nonlinear variant of Raman spectroscopy. The basic concepts of Raman and CARS are introduced first, and subsequent biomedical applications of Raman and CARS are described. Raman microspectroscopy is applied to both in-vivo and in-vitro tissue diagnostics, and the characterization and identification of individual mammalian cells. These applications benefit from the fact that Raman spectra provide specific information on the chemical composition and molecular structure in a label-free and nondestructive manner. Combining the chemical specificity of Raman spectroscopy with the spatial resolution of an optical microscope allows recording hyperspectral images with molecular contrast. We also elaborate on interfacing Raman spectroscopic tools with other technologies such as optical tweezing, microfluidics and fiber optic probes. Thereby, we aim at presenting a guide into one exciting branch of modern biophotonics research.
Topics: Animals; Colon; Diagnostic Imaging; Histological Techniques; Lipids; Mice; Microscopy; Microspectrophotometry; Optics and Photonics; Sensitivity and Specificity; Spectrum Analysis, Raman
PubMed: 22559673
DOI: 10.1117/1.JBO.17.4.040801 -
The Journal of General Physiology Apr 1972Larvae of the mosquito Aedes aegypti have a cluster of four ocelli on each side of the head. The visual pigment of each ocellus of mosquitoes reared in darkness was...
Larvae of the mosquito Aedes aegypti have a cluster of four ocelli on each side of the head. The visual pigment of each ocellus of mosquitoes reared in darkness was characterized by microspectrophotometry, and found to be the same. Larval mosquito rhodopsin (lambda(max) = 515 nm) upon short irradiation bleaches to a stable photoequilibrium with metarhodopsin (lambda(max) = 480 nm). On long irradiation of glutaraldehyde-fixed tissues or in the presence of potassium borohydride, bleaching goes further, and potassium borohydride reduces the product, retinal, to retinol (vitamin A(1)). In the presence of hydroxylamine, the rhodopsin bleaches rapidly, with conversion of the chromophore to retinaldehyde oxime (lambda(max) about 365 nm).
Topics: Aedes; Animals; Borohydrides; Darkness; Hydroxylamines; Larva; Microscopy; Potassium; Retinal Pigments; Spectrophotometry; Ultraviolet Rays; Vitamin A
PubMed: 5029551
DOI: 10.1085/jgp.59.4.401 -
The Journal of Adhesive Dentistry Feb 2012The biodegradation of exposed dentin collagen within the adhesive/dentin (a/d) interface is one of the main reasons for composite restoration failures and seriously...
PURPOSE
The biodegradation of exposed dentin collagen within the adhesive/dentin (a/d) interface is one of the main reasons for composite restoration failures and seriously affects the durability of dental restorations. In the present study, the objective was to investigate whether the inclusion of the cross-linking reagent (glutaraldehyde, GA) in the adhesive would increase collagen biodegradation resistance within the a/d interface.
MATERIALS AND METHODS
The model adhesive consisted of ~60 % monomers (HEMA/bis-GMA, 45/55 wt/wt) and ~ 40 % ethanol as a solvent. 5% GA was added to the above formulation. After the dentin surfaces were etched for 15 s with 35% phosphoric acid, rinsed with water and blotted dry, adhesives both with and without GA were applied and polymerized by visible light for 20 s. These a/d specimens were immersed in the biodegradation solution (prepared by adding 160 mg collagenase in 1 liter of TESCA buffer solution) for up to 30 days after proceeding with the sectioning/fracture to expose the a/d interfaces. The specimens were analyzed using SEM and micro-Raman spectroscopy.
RESULTS
SEM results indicated that for the adhesive without GA, there were many voids and a loss of collagen fibrils in the a/d interface after being challenged by the biodegradation solution. The Raman spectra collected from the interface showed that the amide I of collagen at 1667 cm-1 obviously decreased, indicating a removal of collagen fibrils during the degradation process. For the adhesive containing GA, the collagen fibrils within the interface did not degrade at all, which was also confirmed by the Raman results.
CONCLUSION
The results corroborate the previous findings that by using the current adhesive system and wet bonding, the collagen fibrils in the a/d interface are largely unprotected and easily undergo biodegradation. Directly including cross-linking agents in the adhesive could protect collagen fibrils from degradation in situ within the a/d interface.
Topics: Acid Etching, Dental; Bisphenol A-Glycidyl Methacrylate; Collagen; Collagenases; Cross-Linking Reagents; Dental Bonding; Dentin; Ethanol; Glutaral; Humans; Light-Curing of Dental Adhesives; Materials Testing; Methacrylates; Microscopy, Electron, Scanning; Microspectrophotometry; Phosphoric Acids; Polymerization; Resin Cements; Solvents; Spectrum Analysis, Raman; Surface Properties; Time Factors
PubMed: 21594232
DOI: 10.3290/j.jad.a21494 -
The Journal of Neuroscience : the... Dec 1999Color discrimination requires the input of different photoreceptor cells that are sensitive to different wavelengths of light. The Drosophila visual system contains...
Blue- and green-absorbing visual pigments of Drosophila: ectopic expression and physiological characterization of the R8 photoreceptor cell-specific Rh5 and Rh6 rhodopsins.
Color discrimination requires the input of different photoreceptor cells that are sensitive to different wavelengths of light. The Drosophila visual system contains multiple classes of photoreceptor cells that differ in anatomical location, synaptic connections, and spectral sensitivity. The Rh5 and Rh6 opsins are expressed in nonoverlapping sets of R8 cells and are the only Drosophila visual pigments that remain uncharacterized. In this study, we ectopically expressed Rh5 and Rh6 in the major class of photoreceptor cells (R1-R6) and show them to be biologically active in their new environment. The expression of either Rh5 or Rh6 in "blind" ninaE(17) mutant flies, which lack the gene encoding the visual pigment of the R1-R6 cells, fully rescues the light response. Electrophysiological analysis showed that the maximal spectral sensitivity of the R1-R6 cells is shifted to 437 or 508 nm when Rh5 or Rh6, respectively, is expressed in these cells. These spectral sensitivities are in excellent agreement with intracellular recordings of the R8p and R8y cells measured in Calliphora and Musca. Spectrophotometric analyses of Rh5 and Rh6 in vivo by microspectrophotometry, and of detergent-extracted pigments in vitro, showed that Rh5 is reversibly photoconverted to a stable metarhodopsin (lambda(max) = 494 nm), whereas Rh6 appears to be photoconverted to a metarhodopsin (lambda(max) = 468 nm) that is less thermally stable. Phylogenetically, Rh5 belongs to a group of short-wavelength-absorbing invertebrate visual pigments, whereas Rh6 is related to a group of long-wavelength-absorbing pigments and is the first member of this class to be functionally characterized.
Topics: Amino Acid Sequence; Animals; Animals, Genetically Modified; Color Perception; Drosophila melanogaster; Invertebrates; Photochemistry; Photoreceptor Cells, Invertebrate; Phylogeny; Retinal Pigments; Rhodopsin; Spectrum Analysis
PubMed: 10594055
DOI: 10.1523/JNEUROSCI.19-24-10716.1999