functions in cell biology
CURRENT FRONTIERS AND PERSPECTIVES IN CELL BIOLOGY doc
... claudins and the TAMP proteins Integral proteins establish cell- cell contact in the intercellular space The peripheral proteins, act as a bridge between the integral proteins and the actin cytoskeleton ... types of cell adhesion molecules such as integrins (e.g endothelial JAM-A with integrin LFA-1 in leukocytes and integrin ανβ3 in endothelial cells; endothelial JAM-B with integrin VLA4 in Tight ... paracingulin JACOP and paracingulin are the same protein JACOP/paracingulin is a 160 kDa protein with a domain organization similar to cingulin and 40% sequence identity JACOP/paracingulin besides...
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Atomic Force Microscopy in Cell Biology_Methods in Cell Biology Volume 68 pptx
... coordinate of unfolding is imposed by the direction of pulling, and in this way unfolding events occurring in a single protein can be examined Each peak is attributed to a breakage of a main stabilizing ... differences in the binding affinities of proteins to surfaces In addition, in vitro cell cultures require cell- surface adhesion, which is mediated by a sublayer of adsorbed proteins Protein adsorption ... for instance, to membrane and other molecular structures for a certain time and to track their motion A Mechanics of Molecular Motors Kinesins and kinesin-like proteins are of wide interest in biology...
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Atomic Force Microscopy in Cell Biology ppt
... coordinate of unfolding is imposed by the direction of pulling, and in this way unfolding events occurring in a single protein can be examined Each peak is attributed to a breakage of a main stabilizing ... differences in the binding affinities of proteins to surfaces In addition, in vitro cell cultures require cell- surface adhesion, which is mediated by a sublayer of adsorbed proteins Protein adsorption ... for instance, to membrane and other molecular structures for a certain time and to track their motion A Mechanics of Molecular Motors Kinesins and kinesin-like proteins are of wide interest in biology...
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Atomic Force Microscopy in Cell Biology Episode 1 Part 1 ppt
... data points per line, obtaining at the end a square grid of data points each corresponding to the relative x, y, and z coordinates in space of the sample surface (11) Usually during scanning data ... of curvature, maintaining reproducibility and durability (8) The cantilever carrying the tip is attached to a small glass “chip” that allows easy handling and positioning in the instrument There ... cantilever mounting 5.2 Imaging in Liquid One of the main reasons for the success of AFM in biomedical investigations is its ability to scan samples in physiological condition, that is, immersed in liquid...
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Atomic Force Microscopy in Cell Biology Episode 1 Part 2 pptx
... mathematically modeling the nonlinear behavior of the scanner, finding the parameters for a correction algorithm imaging a known grid, and then applying the algorithm during scanning using the parameters ... amplitude and dampening (in AC modes), feedback gain (sometimes separated into a proportional gain setting and integral-derivative setting), low pass filters, scan speed, and so on The setting of these ... depending on the mode of operation, have been investigated and explanations have been proposed (10–14) Because we are now interested in showing a general overview of the subject for beginners in...
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Atomic Force Microscopy in Cell Biology Episode 1 Part 3 pdf
... 180 min) The specific binding signal quickly builds up and remains stable The interaction is fully reversible and can be broken by shifting the equilibrium of the binding reaction by injecting ... coated by using unspecifically interacting proteins (e.g., bovine serum albumin) In protein detection experiments, larger fluctuations of the cantilevers are observed (e.g., Fig 7) than in the ssDNA–ssDNA ... of Cell Dynamics As mentioned in Subheading 1., the AFM is uniquely capable of tracking the temporal evolution of systems consisting of living cells in a biocompatible fluid There are distinctly...
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Atomic Force Microscopy in Cell Biology Episode 1 Part 4 potx
... process increases the unevenness of the sample surface thus increasing contrast In addition, information concerning the sample’s molecular composition can be revealed and help in the interpretation ... embedded in a matrix of proteoglycans, and keratocytes The former are interspersed between the lamellae, forming an interlinking network throughout the cornea (11–13) AFM has been recently introduced ... fibroblasts in vitro by atomic force microscopy Cytometry 36, 254–264 Pietrasanta, L I., Schaper, A., and Jovin, T M (1994) Imaging subcellular structures of rat mammary carcinoma cells by scanning force...
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Atomic Force Microscopy in Cell Biology Episode 1 Part 5 doc
... µm in (A), µm in (B), and 0.5 µm in (C) 90 Takano and Abe 11 Immerse the samples in 3-butyl-ethyl acetate solution for 15 12 Put a sample in a cage and perform critical point drying 3.2 Scanning ... The program opens the input text file and examines the first line consisting of 500 data points of height values recorded in nanometres The line of data is then derivatized using a first-order algorithm ... the slide glass with spermatozoa in turn to 80 (5 min), 90 (5 min), 95 (5 min), and 100% (10 min, twice) ethyl alcohol solutions for dehydration Figs 3–6 (facing page) AFM images of mouse epididymal...
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Atomic Force Microscopy in Cell Biology Episode 1 Part 6 pdf
... resulted in diminished cell motion and improved the study Wellresolved topographic information could be obtained Zooming -in allows the recording of pictures with increasing detail Especially in fluid ... developing or restoring limb form through the process of endochondral bone formation In the beginning, mesenchymal cells condense and cells in the core differentiate into chondrocytes, and the cells ... done in honey or highly viscous material Tip contamination often occurs This is explained in detail in Fig 11 7.9 Cells Can Contaminate or Stick to the Scanning Tip Occasionally during the scanning...
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Atomic Force Microscopy in Cell Biology Episode 1 Part 7 pot
... dentin (9–13) 1.2 Continuous and Sequential Longitudinal Studies and Reference Layers Continuous scanning is conducted by obtaining images continuously during demineralization in the wet cell ... imaging Standard silicon nitride tips are quite sufficient for good imaging in contact mode on living cells as the visco-elasticity of the cell is the main limiting factor in resolution: in fact, ... AFM 135 nominal setpoint used during imaging in the case of curve 3, we have an indentation of 950 nm, and for curve we find 200 nm We need to keep these features and figures in mind in order to...
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Atomic Force Microscopy in Cell Biology Episode 1 Part 8 pptx
... surrounding intertubular dentin, leaving enlarged tubule lumens (Fig 4B) Exposure increments are continued in steps of 10 s to a minute or more to follow continued changes in the dentin structure ... considerable interest in measuring dentin or other calcified tissues as modified by demineralization or hypomineralization For example, etching procedures are used widely for bonding dentin; dentin caries ... penetrate into the dentin and fill the tubules This becomes evident during demineralization as the filled tubules become increasingly obvious as surrounding dentin recedes leaving them sticking up...
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Atomic Force Microscopy in Cell Biology Episode 1 Part 9 docx
... sarcolemmal structure, including the glycocalyx, are preserved in these isolated cells The structure of intracellular membranes and organelles also shows excellent preservation, indistinguishable from ... laterally into a cell commonly led to destruction of the probe Vibrational/acoustic shielding was achieved by mounting the microscope in a PicoIsolation chamber (Molecular Imaging Co) during scanning ... appropriate instrument settings depending on the imaging mode selected (contact, intermittent contact, noncontact; see Note 7) In general, accurate feedback setting is necessary to obtain the maximum...
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Atomic Force Microscopy in Cell Biology Episode 1 Part 10 potx
... monitor allowing easy positioning of the cell of interest under the tip and real time observation of the AFM scanning process In this AFM Imaging of Living Cells 205 For living cells, a low resonance ... Imaging of Living Cells 201 15 Imaging Surface and Submembranous Structures in Living Cells With the Atomic Force Microscope Notes and Tricks Filip Braet and Eddie Wisse Introduction In 1986, Binnig ... microscope Nonviable cells stain blue and viable cells exclude the stain (11) 3.5 Imaging and Image Interpretation During Scanning 3.5.1 Z-Height Limitations AFM imaging can result in artifactual images...
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Atomic Force Microscopy in Cell Biology Episode 2 Part 1 ppsx
... obtained with a loading force of nN showing fine membranous detail in the form of microvilli (>) Cell border (*), 20 µm × 20 µm (C) Increasing the loading force with a factor 10 resulted in the ... understanding of both microscopies, providing a limit of confidence for AFM imaging of living cells Finally, the use of a video camera installed on the eyepieces of the inverted microscope in combination ... be kept in the interval of 0.8–0.9 for air imaging and 0.90–0.95 for imaging in liquid Scan rates in tapping mode are relatively low (1–3 Hz, depending on the field size) For imaging individual...
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Atomic Force Microscopy in Cell Biology Episode 2 Part 2 ppt
... proteins inserted into lipid films by AFM in air because such proteins interfere Imaging Structures of Surfactant Films 233 with the lipid packing (19) In case of dysfunction of surfactant as in ... online mode, and allow the tip to come in contact with the films surface Set initial scan parameters to Integral Gain–2 and Proportional Gain–3, beginning with the settings recommended for the instrument ... increasing surface packing density driven by increasing lateral pressure (11) The inherent changes of packing the lipid in a film undergoing lateral phase separation allows for the imaging of...
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Atomic Force Microscopy in Cell Biology Episode 2 Part 3 pps
... followed by rinsing in deionized water and air drying Stain the slides in 5% Giemsa solution at room temperature for 45 min, rinse with water, and allow to dry 3.4 Protein Digestion After washing three ... high in order to avoid salt solution creeping into the piezos of the AFM head (see manual for details) 3.4 Single -Cell Volume Measurement in a Living Cell Bring the cantilever close to the cell ... both to calculate the loading force of the past experiment 3.5 Single -Cell Volume Measurement in a Nonliving, Fixed Cell In addition to cell volume measurements on living cells, it is also possible...
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Atomic Force Microscopy in Cell Biology Episode 2 Part 4 docx
... (YPYDVPDYA) epitope tags into the amino acid sequence of the extracellular domains of ENaC subunits The nine amino acids that are replaced in and subunits are indicated in bold For insertion of the ... have been generated against intracellular (N and/or C termini) epitopes of the subunits Insertion of epitope tags into the extracellular domains of transmembrane proteins is a widely used approach ... Force Mapping (31) allows a quantification of the unindented cell volume, independent of the loading force and independent of the elastic modulus of the cell The following section explains the...
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Atomic Force Microscopy in Cell Biology Episode 1 Part 1 pdf
... Imaging of Intact Cells Imaging of Cells Imaging of Isolated Membranes Conclusion and Perspectives References 52 52 53 53 56 63 63 64 v vi Contents Measuring the Elastic Properties of Living Cells ... 93106 Peter Hinterdorfer (115), Institute for Biophysics, University of Linz, A-4040 Linz, Austria J K Heinrich Horber (1), Cell Biology and Biophysics Program, European Molecular ¨ Biology Laboratory, ... touching things is an essential prerequisite to manipulating things, and the ability to feel and to manipulate single molecules and atoms certainly marks another of these revolutionizing steps in...
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Atomic Force Microscopy in Cell Biology Episode 1 Part 2 pps
... coordinate of unfolding is imposed by the direction of pulling, and in this way unfolding events occurring in a single protein can be examined Each peak is attributed to a breakage of a main stabilizing ... differences in the binding affinities of proteins to surfaces In addition, in vitro cell cultures require cell- surface adhesion, which is mediated by a sublayer of adsorbed proteins Protein adsorption ... for instance, to membrane and other molecular structures for a certain time and to track their motion A Mechanics of Molecular Motors Kinesins and kinesin-like proteins are of wide interest in biology...
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