Contacts of chick fibroblasts on glass: results and limitations of quantitative interferometry

1988 ◽  
Vol 90 (2) ◽  
pp. 215-224
Author(s):  
J. Bailey ◽  
D. Gingell
Keyword(s):  
Refractive Index ◽  
Cell Surface ◽  
Limb Bud ◽  
Membrane Bilayer ◽  
Focal Contacts ◽  
Wide Range ◽  
Chick Heart ◽  
Close Contacts ◽  
Migrating Cells

We have examined the contacts made by explanted chick heart and limb bud fibroblasts after 24–48 h on glass, using quantitative interference reflection microscopy (IRM). Contacts beneath very thin cytoplasmic lamellae were avoided because the images of such contacts depend on the thickness of the lamellae. Plaque-like focal contacts, distinguished on the basis of shape and low irradiance (darkness), are intimate adhesions to the substratum. These images can be interpreted if it is assumed that microfilaments associated with the lower membrane increase the local cytoplasmic refractive index. The range of irradiances measured for focal contacts was found to be rather wide, and our modelling shows that the most likely explanation for this is that the images receive variable contributions from the adjacent cytoskeleton. For this reason it is particularly difficult to assign a characteristic thickness for these contacts from IRM data. Close contacts, seen principally as ‘grey’ regions under migrating cells at the edges of the explants, also show a wide range of irradiances. Unlike focal contacts, it is not necessary to postulate any involvement of the cytoskeleton in their images and they can be modelled as regions where an aqueous glycocalyx zone about 20–30 nm thick separates the membrane bilayer from the glass. Paler grey regions that also look like close contacts are apparently formed where the cell surface has lifted several tens of nanometres from the glass.

Rapid cellular translocation is related to close contacts formed between various cultured cells and their substrata

1982 ◽  
Vol 54 (1) ◽  
pp. 23-34
Author(s):  
J. Kolega ◽  
M.S. Shure ◽  
W.T. Chen ◽  
N.D. Young
Keyword(s):  
Cultured Cells ◽  
Mdck Cells ◽  
Time Lapse ◽  
Cell Types ◽  
Epidermal Cells ◽  
Cellular Translocation ◽  
Focal Contacts ◽  
Contact Patterns ◽  
Chick Heart ◽  
Close Contacts

Interference-reflection microscopy combined with time-lapse cinemicrography was used to examine the relationship between cell-to-substratum contact patterns and the speeds of translocation for a variety of cell types. Rapid translocation of amphibian leukocytes (average speed = 9.0 micron/min), amphibian epidermal cells (7 micron/min) and teleost epidermal cells (7 micron/min) was found to correlate with patterns of broad grey close contacts. Similar contact patterns were found under freshly seeded (2 h) chick heart fibroblasts (moving 1–3 micron/min), the rapidly advancing (1-5 micron/min) margin of spreading human WI-38 fibroblasts, and isolated MDCK canine epithelial cells (0.5-1.0 micron/min). Conversely, numerous dark streaks of focal contact were found associated with the slow rate of translocation displayed by older cultures (72 h) of chick fibroblasts (less than 0.1 micron/min), well-spread WI-38 cells (less than or equal to 0.3 micron/min) and confluent MDCK cells (less than 0.01 micron/min). It is concluded that close contacts, but not focal contacts, are associated with rapid cellular translocation, and that the build-up of focal contacts is associated with reduced cellular translocation and maintenance of the spread cell shape.

Simplified coupling power model for fibers fusion

2009 ◽  
Vol 17 (3) ◽  
Author(s):  
J. Saktioto ◽  
J. Ali ◽  
M. Fadhali
Keyword(s):  
Refractive Index ◽  
Propagation Constant ◽  
Tunable Filter ◽  
Experimental Result ◽  
Total Power ◽  
Optical Parameters ◽  
Coupling Region ◽  
Ratio Distribution ◽  
Hydrogen Flow ◽  
Wide Range

AbstractFiber coupler fabrication used for an optical waveguide requires lossless power for an optimal application. The previous research coupled fibers were successfully fabricated by injecting hydrogen flow at 1 bar and fused slightly by unstable torch flame in the range of 800–1350°C. Optical parameters may vary significantly over wide range physical properties. Coupling coefficient and refractive index are estimated from the experimental result of the coupling ratio distribution from 1% to 75%. The change of geometrical fiber affects the normalized frequency V even for single mode fibers. V is derived and some parametric variations are performed on the left and right hand side of the coupling region. A partial power is modelled and derived using V, normalized lateral phase constant u, and normalized lateral attenuation constant, w through the second kind of modified Bessel function of the l order, which obeys the normal mode and normalized propagation constant b. Total power is maintained constant in order to comply with the energy conservation law. The power is integrated through V, u, and w over the pulling length of 7500 µm for 1-D. The core radius of a fiber significantly affects V and power partially at coupling region rather than wavelength and refractive index of core and cladding. This model has power phenomena in transmission and reflection for an optical switch and tunable filter.

An analysis of chick limb bud intercellular adhesion underlying the establishment of cartilage aggregates in suspension culture

1990 ◽  
Vol 96 (3) ◽  
pp. 527-536
Author(s):  
J.A. Bee ◽  
K. von der Mark
Keyword(s):  
Cell Surface ◽  
Cell Aggregation ◽  
Intercellular Adhesion ◽  
Limb Bud ◽  
Affinity Column ◽  
Type I ◽  
Surface Plasma ◽  
Aggregation Mechanism ◽  
Fab Fragments ◽  
Exogenous Calcium

To examine the mechanism of intercellular adhesion in the establishment of limb skeletal elements we have investigated the process of limb bud cell aggregation in vitro. Limb bud cells are aggregation-competent immediately after their trypsin:collagenase dissociation in the absence of calcium. This aggregation is largely Ca2(+)-independent (CI) and is completely and reversibly inhibited by cycloheximide. In contrast, when limb bud cells are first allowed to recover from Ca2(+)-free trypsin:collagenase dissociation, aggregation of the surviving population is exclusively Ca2(+)-dependent (CD) and completely and reversibly inhibited by cycloheximide. The presence of exogenous calcium during initial cell dissociation retains a functional CD aggregation mechanism. However, incubation of such cells with EGTA releases the CD component and converts the cells to a predominantly CI aggregation. Rabbits were immunized with limb bud cells exhibiting the recovered CD aggregation mechanism and the resulting immune sera were screened for their effect on cell aggregation. Relative to pre-immune sera, intact immune IgG agglutinated dissociated limb bud cells whilst immune Fab fragments inhibited their aggregation. The aggregation-inhibiting antiserum recognizes five major limb bud cell surface components with apparent molecular weights of 72K, 50K, 23K, 14.5K and 8.5K (K = 10(3) Mr), respectively. Limb bud cell surface plasma membranes were isolated by sucrose gradient density centrifugation and detergent-solubilized proteins coupled to Sepharose 4B with cyanogen bromide. Equivalent cell surface plasma membrane proteins were 125I-iodinated and applied to the affinity column. Limb bud cell surface protein affinity chromatography in the presence of exogenous calcium yields a single protein with an apparent molecular weight of approximately 8.5 K. This protein molecule elutes at 0.6 M NaCl, indicating a high affinity, is recognized by the aggregation-inhibiting antiserum, and is itself capable of inhibiting CD limb bud cell aggregation. Fab fragments prepared from rabbit antisera specifically directed against the affinity-purified material also inhibit CD limb bud cell aggregation and this inhibition is neutralized by the 8.5 K protein. Our data thus demonstrate that CD limb bud cell aggregation is not mediated by fibronectin and/or collagen type I and indicate that this process is governed by a novel 8.5 K cell adhesion molecule.

Functional partitioning of beta1 integrins revealed by activating and inhibitory mAbs

1997 ◽  
Vol 110 (21) ◽  
pp. 2647-2659 ◽  
Author(s):  
M.T. Cruz ◽  
C.L. Dalgard ◽  
M.J. Ignatius
Keyword(s):  
Cell Surface ◽  
Actin Cytoskeleton ◽  
Dermal Fibroblasts ◽  
The Other ◽  
Embryonic Chick ◽  
Cell Surfaces ◽  
Double Labeling ◽  
Focal Contacts ◽  
Functional Partitioning ◽  
Discrete Populations

Integrins exist in different activation states on the surfaces of cells. Addition of the proper signal, ligand, or antibody can alter the activation state of these molecules. We report here the identification of two immunocytochemically distinct populations of beta1 integrins on fixed embryonic chick dermal fibroblasts. One population, recognized by the integrin activating mAb TASC, localizes to discrete regions of the cell, most likely focal contacts. These integrins co-localize with other proteins, such as vinculin and F-actin, and their retention at these sites is dependent on the actin cytoskeleton. The other population, identified with the inhibitory mAb W1B10, is more evenly distributed throughout the cell surface, and its pattern remains unchanged after disruption of the actin cytoskeleton. Double labeling experiments using Fab fragments of TASC alongside whole W1B10 IgG revealed non-overlapping staining patterns. These results show that it is possible to visualize and study discrete populations of integrins on cell surfaces using two different antibodies. We hypothesize that these antibodies report differences in the distribution of receptors in two different states. A model is proposed describing the ligand independent recruitment of integrins based on these findings and results from other labs.

scholarly journals Glucosylceramide Plays a Role in Fungal Germination, Lipid Raft Organization and Biofilm Adhesion of the Pathogenic Fungus Scedosporium aurantiacum

2020 ◽  
Vol 6 (4) ◽  
pp. 345
Author(s):  
Victor Pereira Rochetti ◽  
Rodrigo Rollin-Pinheiro ◽  
Evely Bertulino de Oliveira ◽  
Mariana Ingrid Dutra da Silva Xisto ◽  
Eliana Barreto-Bergter
Keyword(s):  
Cell Surface ◽  
Pathogenic Fungus ◽  
Clinical Manifestations ◽  
Tube Length ◽  
Total Biomass ◽  
Fungal Cell ◽  
Germ Tube Length ◽  
Wide Range ◽  
Fluorescent Stain ◽  
Altered Lipid

Infections caused by Scedosporium species present a wide range of clinical manifestations, from superficial to disseminated, especially in immunocompromised patients. Glucosylceramides (GlcCer) are glycosphingolipids found on the fungal cell surface and play an important role in growth and pathogenicity processes in different fungi. The present study aimed to evaluate the structure of GlcCer and its role during growth in two S. aurantiacum isolates. Purified GlcCer from both isolates were obtained and its chemical structure identified by mass spectrometry. Using ELISA and immunofluorescence techniques it was observed that germination and NaOH-treatment of conidia favor GlcCer exposure. Monoclonal anti-GlcCer antibody reduced germination when cultivated with the inhibitor of melanin synthesis tricyclazole and also reduced germ tube length of conidia, both cultivated or not with tricyclazole. It was also demonstrated that anti-GlcCer altered lipid rafts organization, as shown by using the fluorescent stain filipin, but did not affect the susceptibility of the cell surface to damaging agents. Anti-GlcCer reduced total biomass and viability in biofilms formed on polystyrene plates. In the presence of anti-GlcCer, germinated S. aurantiacum conidia and biofilms could not adhere to polystyrene with the same efficacy as control cells. These results highlight the relevance of GlcCer in growth processes of S. aurantiacum.

scholarly journals Improved Refractive Index-Sensing Performance of Multimode Fano-Resonance-Based Metal-Insulator-Metal Nanostructures

Nanomaterials ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 2097
Author(s):  
Yuan-Fong Chou Chau ◽  
Chung-Ting Chou Chao ◽  
Siti Zubaidah Binti Haji Jumat ◽  
Muhammad Raziq Rahimi Kooh ◽  
Roshan Thotagamuge ◽  
...  
Keyword(s):  
Refractive Index ◽  
Plasmon Resonance ◽  
Fano Resonance ◽  
High Sensitivity ◽  
Metal Nanostructures ◽  
Refractive Index Sensor ◽  
Metal Insulator ◽  
Wide Range ◽  
Metal Insulator Metal ◽  
Mode 2

This work proposed a multiple mode Fano resonance-based refractive index sensor with high sensitivity that is a rarely investigated structure. The designed device consists of a metal–insulator–metal (MIM) waveguide with two rectangular stubs side-coupled with an elliptical resonator embedded with an air path in the resonator and several metal defects set in the bus waveguide. We systematically studied three types of sensor structures employing the finite element method. Results show that the surface plasmon mode’s splitting is affected by the geometry of the sensor. We found that the transmittance dips and peaks can dramatically change by adding the dual air stubs, and the light–matter interaction can effectively enhance by embedding an air path in the resonator and the metal defects in the bus waveguide. The double air stubs and an air path contribute to the cavity plasmon resonance, and the metal defects facilitate the gap plasmon resonance in the proposed plasmonic sensor, resulting in remarkable characteristics compared with those of plasmonic sensors. The high sensitivity of 2600 nm/RIU and 1200 nm/RIU can simultaneously achieve in mode 1 and mode 2 of the proposed type 3 structure, which considerably raises the sensitivity by 216.67% for mode 1 and 133.33% for mode 2 compared to its regular counterpart, i.e., type 2 structure. The designed sensing structure can detect the material’s refractive index in a wide range of gas, liquids, and biomaterials (e.g., hemoglobin concentration).

Wide-range tunable, dual-band, background refractive index insensitive terahertz absorber based on graphene and Dirac semimetal

2021 ◽  
Vol 60 (02) ◽  
Author(s):  
Shuaizhao Wang ◽  
Ronghui Xu ◽  
Zujun Qin ◽  
Houquan Liu ◽  
Shijie Deng ◽  
...  
Keyword(s):  
Refractive Index ◽  
Dual Band ◽  
Terahertz Absorber ◽  
Dirac Semimetal ◽  
Wide Range

The analysis of spatial distributions in mixed cell populations: a statistical method for detecting sorting out

Development ◽  
1971 ◽  
Vol 26 (1) ◽  
pp. 135-156
Author(s):  
R. A. Elton ◽  
C. A. Tickle
Keyword(s):  
Plant Communities ◽  
Quantitative Measure ◽  
Cell Types ◽  
Limb Bud ◽  
Cell Populations ◽  
Spatial Distributions ◽  
Mixed Cell ◽  
Mixed Aggregates ◽  
Chick Heart ◽  
Gyratory Shaker

1. This work presents a quantitative measure, α, of the degree of segregation of two cell types in sections of aggregates, and some results obtained with the measure relating to ‘sorting out’. The method is designed particularly for the case where labelling of one type of cell is incomplete, and the importance of this effect is assessed. Possible problems in formulating such a model are discussed. The measure α is compared with methods used in investigations of segregation in plant communities. 2. Segregation of chick heart and limb-bud cells in mixed aggregates has been analysed using α. In control aggregates of mixtures of labelled and unlabelled cells of one type, α is near to its random value of 1, and we suggest that the departure from random can be adequately accounted for by cell division. In mixed aggregates, significant segregation is consistently found, even in aggregates formed after 2 and 4 h. Both disaggregation procedures (EDTA, trypsin or trypsin + EDTA) and reaggregation methods (reciprocating or gyratory shaker) are found to have an effect on the degree of segregation. Possible reasons for these findings are discussed. 3. Positioning of the cells relative to the outside of aggregates is also investigated for some of the aggregates.

Cell death and the development of limb form and skeletal pattern in normal and wingless (ws) chick embryos

Development ◽  
1973 ◽  
Vol 30 (3) ◽  
pp. 753-772
Author(s):  
J. R. Hinchliffe ◽  
D. A. Ede
Keyword(s):  
Cell Death ◽  
Phenotypic Expression ◽  
Mesenchymal Cell ◽  
Limb Bud ◽  
Death Process ◽  
Electron Microscopic ◽  
Wide Range ◽  
Skeletal Pattern ◽  
Autoradiographic Analysis ◽  
Mutant Phenotypes

The wingless condition resulting from the action of the sex-linked wingless (ws) gene arises from the precocious appearance of cell death in the anterior necrotic zone (ANZ) of the forelimb-bud at stage 19 (3 days) and its progressive extension beyond its normal area during stages 20–23. A similar though less pronounced effect occurs in the hindlimb-bud. Although some wingless hindlimb-buds are normal, others are affected by the precocious appearance of cell death in the ANZ. The ws wingless mutant resembles the different wingless mutant investigated by Zwilling (1956) in that the apical ectodermal ridge (AER) is absent in most ws wing-buds. AER absence could be due to ws mesenchymal cell death interfering with the production of apical ectodermal maintenance factor (AEMF), which Zwilling claims is necessary to maintain the AER which plays an essential role in inducing limb outgrowth. Wingless mutant phenotypes range from birds with rudimentary wings and normal legs through a modal type with forelimbs absent and hindlimbs normal to wingless and legless forms showing a high degree of expressivity. Individual wingless embryos vary in the degree to which the precocious ANZ appearing at 3 days is extended into the limb-bud and the wide range of wingless phenotypic expression is attributed to this variation. Electron microscopic and histochemical analysis of the cell death process in wingless wing-buds revealed the presence of both isolated dead cells and macrophages, which contained intense acid phosphatase activity. These findings are interpreted as showing that isolated dead cells are ingested by neighbouring mesenchymal cells which thus become transformed into macrophages, first ingesting and then digesting further dead cells. A study was made of the origin of the anomalous hindlimb condition, including absence or reduction of the tibia and digits, characteristic of severely affected wingless embryos. Autoradiographic analysis of the pattern of 35SO4 uptake revealed that at stage 24/5 (4½ days) wingless hindlimb-buds which were smaller than normal had a normal prospective fibula region, but that the prospective tibia region was small or absent. Thus the effect of a precocious hindlimb ANZ at stages 19–22 is to reduce or delete the pre-axial prospective tibia at stage 24/5.

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