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Modeling Lysis Dynamcis Of Pore Forming Toxins And Determination Of Mechanical Properties Of Soft MaterialsVaidyanathan, M S 11 1900 (has links) (PDF)
Pore forming toxins are known for their ability to efficiently form transmembrane pores which eventually leads to cell lysis. PFTs have potential applications in devel-oping novel drug and gene delivery strategies. Although structural aspects of many pore forming toxins have been studied, very little is known about the dynamics and subsequent rupture mechanisms. In the first part of the thesis, a combined experimental and modeling study to understand the lytic action of Cytolysin A (ClyA) toxins on red blood cells (RBCs) has been presented. Lysis experiments are carried out on a 1% suspension of RBCs for different initial toxin concentrations ranging from 100 – 500 ng/ml and the extent of lysis is monitored spectrophotometrically. Using a mean field approach, we propose a non – equilibrium adsorption-reaction model to quantify the rate of pore formation on the cell surface. By analysing the model in a pre-lysis regime, the number of pores per RBC to initiate rupture was found to lie between 400 and 800. The time constants for pore formation are estimated to lie between 1-25 s and monomer conformation time scales were found to be 2-4 times greater than the oligomerization times. Using this model, we are able to predict the extent of cell lysis as a function of the initial toxin concentration. Various kinetic models for oligomerization mechanism have been explored. Irreversible sequential kinetic model has the best agreement with the available experimental data. Subsequent to the mean field approach, a population balance model was also formulated.
The mechanics of cell rupture due to pore formation is poorly understood. Efforts to address this issue are concerned with understanding the changes in the membrane mechanical properties such as the modulus and tension in the presence of pores. The second part of the thesis is concerned with using atomic force microscopy to measure the mechanical properties of cells. We explore the possibility of employing tapping mode AFM (TM-AFM) to obtain the elastic modulus of soft samples. The dynamics of TM-AFM is modelled to predict the elastic modulus of soft samples, and predict optimal cantilever stiffness for soft biological samples. From experiments using TM-AFM on Nylon-6,6 the elastic modulus is predicted to lie between 2 and 5 GPa. For materials having elastic moduli in the range of 1– 20 GPa, the cantilever stiffness from simulations is found to lie in the range of 1 – 50 N/m. For soft biological samples, whose elastic moduli are in the range of 10-1000 kPa, a narrower range of cantilever stiffness (0.1 – 0.6 N/m), should be used.
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Beitrag zur Berechnung von Haftkräften auf rauen Oberflächen am Beispiel keramischer Filtersysteme bei der MetallschmelzefiltrationDitscherlein, Lisa 01 December 2021 (has links)
Diese Arbeit setzt sich mit der Messung und Modellierung von Haftkräften auf rauen Oberflächen auseinander, wobei Filtermaterialien aus dem Sonderforschungsbereich 920 mittels Rasterkraftmikroskopie-Methoden untersucht wurden. Durch eine Vielzahl an Haftkraftmessungen konnte gezeigt werden, dass für schlecht benetzende Oberflächen die Rauheit zur Erhöhung der attraktiven Kräfte führt, da vermehrt mit Kapillarwechselwirkungen zu rechnen ist. Aus diesem Grund erfolgte eine Charakterisierung der Oberflächenmorphologien der Filtermaterialproben und eine kritische Bewertung von Modellen, welche aus Kontaktwinkeldaten die Oberflächenenergien ermitteln. Im Modellierungsteil der Arbeit wurden gängige van der Waals-Kraftmodelle mit Berücksichtigung von Retardationseffekten untersucht. Weiterhin wurden AFM-Scans mit dem Schichtmodell von Dagastine verbunden sowie für die Bewertung von Kapillarkraftmodellen verwendet.:Abbildungsverzeichnis
Tabellenverzeichnis
1 Einleitung
2 Stand der Technik
2.1 Reinigung von Metallschmelzen
2.1.1 Stellung des Sonderforschungsbereichs 920 innerhalb der Thematik
der Metallschmelzereinigung
2.1.2 Inklusionen in Metallschmelzen - Schmelzprozess, relevante
Einschlusstypen und Problematik verunreinigter Gussprodukte
2.1.3 Möglichkeiten der Metallschmelzereinigung
2.1.4 Metallschmelzefiltration
2.2 Rasterkraftmikroskopie
2.2.1 Funktionsweise, Kraftspektroskopie und Imaging
2.2.2 Colloidal Probe Technik
2.3 Benetzung und deren Phänomene
2.3.1 Ansätze zur Quantifizierung von Benetzungsgrößen
2.3.2 Diskussion bezüglich der Oberflächenenergiemodelle
2.3.3 Realsystem: Benetzungsphänomene bei Metallschmelzen auf
Keramiken am Beispiel Aluminium
2.4 Haftkräfte und deren Berechnung
2.4.1 DLVO-Kräfte: van der Waals-Wechselwirkungen
2.4.2 nonDLVO-Kräfte
2.4.3 Nanoblasen und Kapillarkräfte
2.4.4 Haftkräfte unter erhöhten Temperaturen, Sinterung
2.4.5 Haftkräfte auf rauen Oberflächen
2.4.6 Kontaktmechanik
2.5 Genutztes Modellsystem, Abgrenzung der Arbeit
3 Material und Methoden
3.1 Material
3.1.1 Substrate
3.1.2 Partikel
3.1.3 Modellschmelze Wasser, Flüssigkeiten für die Kontaktwinkelmessung
3.2 Methoden
3.2.1 Kontaktwinkelmessgerät G10
3.2.2 Rasterkraftmikroskope XE-100 und UHV750
3.2.3 MatLAB-Skripte
3.2.4 Silanisierung
3.3 Vorbetrachtungen
3.3.1 Berechnung der Hamaker-Konstanten der untersuchten Stoffsysteme
3.3.2 Bedeutung eines einheitlichen Protokolls bei AFM- und KWMessungen
4 Auswertung
4.1 Charakterisierung der untersuchten Oberflächen bezüglich ihrer Rauheit
mithilfe von AFM-Scans
4.1.1 Kontaktmechanik und sinnvolle Wahl der Scan-Größen sowie
Lateral-Auflösung
4.1.2 Höhenprofile (z-Werte) der Filtermaterialien
4.1.3 Rauheitskenngrößen der AFM-Scans
4.1.4 Zusammenfassung
4.2 Kontaktwinkelmessungen und Oberflächenenergieverteilungen
4.2.1 Kontaktwinkelmessungen auf den Filtermaterialien
4.2.2 Oberflächenenergie - Komponentenansätze
4.2.3 Oberflächenenergie - Equation of State
4.2.4 Abschätzen der Hamaker-Konstanten aus der Oberflächenenergie
des Feststoffs
4.2.5 Zusammenfassung
4.3 Ergebnisse Kraftspektroskopie
4.3.1 Experimentelle Ergebnisse - Einflussfaktoren Benetzbarkeit,
Rauheit und Gasübersättigung
4.3.2 Sondergeometrien
4.3.3 Nachbetrachtung zu den Ergebnissen des Modellsystems
4.3.4 HT-Messungen mit dem Rasterkraftmikroskop
4.3.5 Zusammenfassung
5 Modellierung
5.1 Berücksichtigung von Retardation bei gängigen van der Waals-
Kraftmodellen auf rauen Oberflächen in der Mechanischen Verfahrenstechnik
5.2 Van der Waals-Kraft-Modelle für raue Oberflächen zur Beschreibung
der experimentellen Daten
5.3 Dagastines Modell in Kombination mit dem Cooper-Ansatz
5.4 Kapillarkräfte durch Nanoblasen auf rauen Oberflächen
5.4.1 Zusammenfassung
6 Zusammenfassung und Ausblick
6.1 Zusammenfassung
6.2 Ausblick
7 Anhang
7.1 Ergänzungen zum Kapitel Stand der Technik
7.2 Ergänzungen zum Kapitel Material und Methoden
7.3 Ergänzungen zum Kapitel Auswertung: Rauheit
7.4 Ergänzungen zum Kapitel Auswertung: Kontaktwinkelmessungen und
Oberflächenenergien
7.5 Ergänzungen zum Kapitel Auswertung: Literatur- und Messdaten
7.6 Ergänzungen zum Kapitel Auswertung: Modellierung
Literaturverzeichnis
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Haftkräfte zwischen technisch rauen OberflächenFritzsche, Jörg 07 October 2016 (has links)
Die eingereichte Dissertation beschäftigt sich mit der messtechnischen Erfassung sowie der Modellierung von Haftkräften zwischen rauen Oberflächen. Dabei wurden durch Variation von Flüssigkeiten sowie dem Nutzen beschichteter Oberflächen verschiedene Benetzungseigenschaften eingestellt und untersucht. Zusätzlich wurden neben dem Kontaktwinkel der untersuchten Systeme die freien Ober- und Grenzflächenenergien bestimmt und mit den Kräften korreliert. Es zeigte sich, dass Haftkräfte auf rauen Oberflächen stets über mehrere Größenordnungen verteilt vorliegen. Die Beschreibung der ermittelten Verteilungen ist dabei entweder durch statistischer Funktionen oder zumindest teils auch durch eine im Rahmen der Arbeit entwickelten Modellierung möglich. Weiterhin zeigte sich, dass eine Unterteilung in verschiedene Haftmechanismen (durch Kapillarbrücken oder van der Waals- sowie polare Wechselwirkungen) vorgenommen werden kann. Kapillarbrücken bilden dabei die größten Kräfte aus. Sie entstehen auf Grund nanoskaliger Blasen (Nanobubbles), welche vor allem auf schlecht benetzenden Oberflächen existieren.
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Single-Molecule Measurements of Complex Molecular Interactions in Membrane Proteins using Atomic Force MicroscopySapra, K. Tanuj 01 March 2007 (has links)
Single-molecule force spectroscopy (SMFS) with atomic force microscope (AFM) has advanced our knowledge of the mechanical aspects of biological processes, and helped us take big strides in the hitherto unexplored areas of protein (un)folding. One such virgin land is that of membrane proteins, where the advent of AFM has not only helped to visualize the difficult to crystallize membrane proteins at the single-molecule level, but also given a new perspective in the understanding of the interplay of molecular interactions involved in the construction of these molecules. My PhD work was tightly focused on exploiting this sensitive technique to decipher the intra- and intermolecular interactions in membrane proteins, using bacteriorhodopsin and bovine rhodopsin as model systems. Using single-molecule unfolding measurements on different bacteriorhodopsin oligomeric assemblies - trimeric, dimeric and monomeric - it was possible to elucidate the contribution of intra- and interhelical interactions in single bacteriorhodopsin molecules. Besides, intriguing insights were obtained into the organization of bacteriorhodopsin as trimers, as deduced from the unfolding pathways of the proteins from different assemblies. Though the unfolding pathways of bacteriorhodopsin from all the assemblies remained the same, the different occurrence probability of these pathways suggested a kinetic stabilization of bacteriorhodopsin from a trimer compared to that existing as a monomer. Unraveling the knot of a complex G-protein coupled receptor, rhodopsin, showed the existence of two structural states, a native, functional state, and a non-native, non-functional state, corresponding to the presence or absence of a highly conserved disulfide bridge, respectively. The molecular interactions in absence of the native disulfide bridge mapped onto the three-dimensional structure of native rhodopsin gave insights into the molecular origin of the neurodegenerative disease retinitis pigmentosa. This presents a novel technique to decipher molecular interactions of a different conformational state of the same molecule in the absence of a high-resolution X-ray crystal structure. Interestingly, the presence of ZnCl2 maintained the integrity of the disulfide bridge and the nature of unfolding intermediates. Moreover, the increased mechanical and thermodynamic stability of rhodopsin with bound zinc ions suggested a plausible role for the bivalent ion in rhodopsin dimerization and consequently signal transduction. Last but not the least, I decided to dig into the mysteries of the real mechanisms of mechanical unfolding with the help of well-chosen single point mutations in bacteriorhodopsin. The monumental work has helped me to solve some key questions regarding the nature of mechanical barriers that constitute the intermediates in the unfolding process. Of particular interest is the determination of altered occurrence probabilities of unfolding pathways in an energy landscape and their correlation to the intramolecular interactions with the help of bioinformatics tools. The kind of work presented here, in my opinion, will not only help us to understand the basic principles of membrane protein (un)folding, but also to manipulate and tune energy landscapes with the help of small molecules, proteins, or mutations, thus opening up new vistas in medicine and pharmacology. It is just a matter of a lot of hard work, some time, and a little bit of luck till we understand the key elements of membrane protein (un)folding and use it to our advantage.
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MECHANOBIOLOGY OF BRAIN-DERIVED CELLS DURING DEVELOPMENTAL STAGESMahajan, Gautam January 2019 (has links)
No description available.
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Development of Chemomechanical Functionalization and Nanografting on Silicon SurfacesLee, Michael Vernon 18 July 2007 (has links) (PDF)
Progress in chemomechanical functionalization was made by investigating the binding of molecules and surface coverage on the silicon surface, demonstrating functionalization of silicon with gases by chemomechanical means, analyzing atomic force microscopy probe tip wear in atomic force microscopy (AFM) chemomechanical nanografting, combining chemomechanical functionalization and nanografting to pattern silicon with an atomic force microscope, and extending chemomechanical nanografting to silicon dioxide. Molecular mechanics of alkenes and alkynes bound to Si(001)-2x1 as a model of chemomechanically functionalized surfaces indicated that complete coverage is energetically favorable and becomes more favorable for longer chain species. Scribing a silicon surface in the presence of ethylene and acetylene demonstrated chemomechanical functionalization with gaseous reagents, which simplifies sample cleanup and adds a range of reagents to those possible for chemomechanical functionalization. Thermal desorption spectroscopy was performed on chemomechanically functionalized samples and demonstrated the similarity in binding of molecules to the scribed silicon surface and to the common Si(001)-2x1 and Si(111)-7x7 surfaces. The wearing of atomic force microscope probe tips during chemomechanical functionalization was investigated by correlating change over time and force with widths of created lines to illustrate the detrimental effect of tip wear on mechanically-driven nanopatterning methods. In order to have a starting surface more stable than hydrogen-terminated silicon, silicon reacted with 1-octene was used as a starting surface for AFM chemomechanical functionalization, producing chemomechanical nanografting. Chemomechanical nanografting was then demonstrated on silicon dioxide using silane molecules; the initial passivating layer reduced the tip friction on the surface to allow only partial nanografting of the silane molecules. These studies broadened the scope and understanding of chemomechanical functionalization and nanografting.
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Préparation et études des propriétés des films magnétiques nanostructures pour des applications en dispositifs magnéto-acoustiques et spintroniques / Preparation and studies of properties of nanostructured magnetic films for applications in magnetoacoustic and spintronic devicesPavlova, Anastasia 08 September 2014 (has links)
Aujourd'hui, les structures basées sur les matériaux ferromagnétiques sont largement utilisées pour différentes applications: mémoires magnéto-résistives à accès non séquentiel, capteurs magnétiques et également nouveaux composants électroniques et dipositifs spintroniques. La tendance générale de l'électronique moderne est une réduction de la dimension des éléments à l'échelle submicronique. Ainsi, les nanostructures magnétiques sont d'un grand intérêt et leurs méthodes de fabrication et propriétés sont étudiées activement.Le but principal de ce travail est la préparation et la recherche expérimentale et théorique des propriétés de nanostructures magnétiques pour applications aux composants magneto-résistifs et phononiques. La lithographie à sonde locale (SPL) et la lithographie par faisceau d’électrons (EBL) ont été utilisées pour la fabrication des nanostructures. De premiers pas ont également été réalisés en fabrication des cristaux phononiques sensibles au champ magnétique. / Nowadays, structures based on ferromagnetic materials are largely used for different applications: random access magneto-resistive memories, magnetic sensors, and also new electronic components and spintronic devices. The general trend of modern electronic is the reduction of dimensions down to submicronic scales. Therefore, the magnetic nanostructures are of great interest and their methods of fabrication and properties largely studied.The main goal of this work is the preparation and experimental and theoretical research on properties of magnetic nanostructures for applications in magnetoresistive and photonic devices. The Scanning Probe Lithography (SPL) and Electron Beam Lithography (EBL) were used for the nanostructures fabrications. First steps were also achieved in fabrication of phononic cristals sensitive the magnetic field.
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Development of Micromachined Probes for Bio-Nano ApplicationsYapici, Murat K. 14 January 2010 (has links)
The most commonly known macro scale probing devices are simply comprised
of metallic leads used for measuring electrical signals. On the other hand,
micromachined probing devices are realized using microfabrication techniques and are
capable of providing very fine, micro/nano scale interaction with matter; along with a
broad range of applications made possible by incorporating MEMS sensing and
actuation techniques. Micromachined probes consist of a well-defined tip structure that
determines the interaction space, and a transduction mechanism that could be used for
sensing a change, imparting external stimuli or manipulating matter.
Several micromachined probes intended for biological and nanotechnology
applications were fabricated, characterized and tested. Probes were developed under two
major categories. The first category consists of Micro Electromagnetic Probes for
biological applications such as single cell, particle, droplet manipulation and neuron
stimulation applications; whereas the second category targets novel Scanning Probe
topologies suitable for direct nanopatterning, variable resolution scanning probe/dip-pen
nanolithography, and biomechanics applications.
The functionality and versatility of micromachined probes for a broad range of
micro and nanotechnology applications is successfully demonstrated throughout the five
different probes/applications that were studied. It is believed that, the unique advantages
of precise positioning capability, confinement of interaction as determined by the probe
tip geometry, and special sensor/actuator mechanisms incorporated through MEMS
technologies will render micromachined probes as indispensable tools for microsystems
and nanotechnology studies.
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Fault detection and diagnosis : application in microelectromechanical systems / Ανίχνευση και διάγνωση σφαλμάτων με εφαρμογές σε μικροηλεκτρομηχανικά συστήματαΡέππα, Βασιλική 07 December 2010 (has links)
This thesis presents the development of a fault detection and diagnosis (FDD) procedure capable of capturing, isolating and identifying multiple abrupt parametric faults. The proposed method relies on parameter estimation deployed in a set membership framework. This approach presupposes the utilization of a linearly parametrizable model and the a priori knowledge of bounded noise errors and parameter perturbations. Under these assumptions, a data-hyperspace is generated at every time instant. The goal of set membership identification (SMI) is the determination of the parametric set, formed as an orthotope or ellipsoid, within which the nominal parameter vector resides and intersects with the data-hyperspace.
The fault detection mechanism is activated when the normal operation of the SMI procedure is interrupted due to an empty intersection of the data-hyperspace and the estimated parametric set. At the detection instant, a resetting procedure is performed in order to compute the parameter set and the data-hyperspace that contain the varied nominal parameter vector, allowing the SMI algorithm to continue its operation. During the fault isolation, consistency tests are executed, relying on the projections of the worst case parametric sets and the ones arisen from the normal operation of SMI. A faulty component is indicated when these projections do not intersect, while the distance of their centers is used for fault identification. In case of the ellipsoidal SMI-based FDD and under the assumption of a time invariant parameter vector, a new fault detection criterion is defined based on the intersection of support orthotopes of ellipsoids. A more accurate estimation of the time instant of fault occurrence is proposed based on the application of a backward-in-time procedure starting from the fault detection instant, while the conditions under which a fault will never be detected by the orthotopic and ellipsoidal SMI based FDD are provided.
This dissertation explores the efficiency of the proposed FDD methodology for capturing failure modes of two microelectromechanical systems; an electrostatic parallel-plate microactuator and a torsionally resonant atomic force microscope. From an engineering point of view, failure modes appeared in the microcomponents of the microactuator and the TR-AFM are encountered as parameter variations and are captured, isolated and identified by the proposed FDD methodology. / Σε αυτή την διατριβή, παρουσιάζεται η ανάπτυξη μιας διαδικασίας Ανίχνευσης και Διάγνωσης Σφαλμάτων, η οποία είναι ικανή να εντοπίζει, απομονώνει και αναγνωρίζει πολλαπλά, απότομα παραμετρικά σφάλματα. H προτεινόμενη μέθοδος βασίζεται στην αναγνώριση του συνόλου συμμετοχής των παραμέτρων. Ο στόχος της Αναγνώρισης Συνόλου Συμμετοχής είναι ο καθορισμός του παραμετρικού συνόλου εντός του οποίου κείται το ονομαστικό διάνυσμα παραμέτρων, δεδομένου ότι το ονομαστικό διάνυσμα παραμέτρων ανήκει επίσης σε έναν υπερχώρο δεδομένων. Το παραμετρικό σύνολο απεικονίζεται ως ένα ορθότοπο ή ένα ελλειψοειδές, λόγω της εύκολης μαθηματικής τους περιγραφής. Έτσι, η διαδικασία Αναγνώρισης Συνόλου Συμμετοχής αντιστοιχεί σε ένα πρόβλημα βελτιστοποίησης, το οποίο αποσκοπεί στον υπολογισμό του ορθοτόπου ή ελλειψοειδούς το οποίο περιέχει το ονομαστικό διάνυσμα παραμέτρων και τέμνεται με τον υπερχώρο δεδομένων.
Ο μηχανισμός Ανίχνευσης Σφαλμάτων ενεργοποιείται όταν διακόπτεται η φυσιολογική λειτουργία της Αναγνώρισης Συνόλου Συμμετοχής, λόγω της κενής τομής μεταξύ των εκτιμώμενου παραμετρικού συνόλου και του υπερχώρου δεδομένων. Τη χρονική στιγμή ανίχνευσης ενός σφάλματος, εφαρμόζεται μια διαδικασία επαναρύθμισης που σκοπεύει στον υπολογισμό του νέου παραμετρικού συνόλου, το οποίο περιέχει το μεταβεβλημένο ονομαστικό διάνυσμα παραμέτρων και τέμνεται με το υπερχώρο δεδομένων. Κατά τη διάρκεια της διαδικασίας απομόνωσης του σφάλματος, εκτελούνται τεστ συμβατότητας, τα οποία βασίζονται στις προβολές των νέων παραμετρικών συνόλων και στις προβολές των παραμετρικών συνόλων χείριστης περίπτωσης, ενώ η απόσταση των κέντρων των προβολών χρησιμοποιείται για αναγνώριση σφάλματος. Στην περίπτωση που η Ανίχνευση και Διάγνωση Σφαλμάτων πραγματοποιείται βασιζόμενη στην Αναγνώριση Συνόλου Συμμετοχής με ελλειψοειδή και θεωρώντας το ονομαστικό διάνυσμα παραμέτρων χρονικά αμετάβλητο, ορίζεται ένα νέο κριτήριο ανίχνευσης σφαλμάτων, χρησιμοποιώντας την τομή των περιβαλλόντων ορθοτόπων των ελλειψοειδών. Σε αυτή την περίπτωση, ένα σφάλμα ανιχνεύεται όταν η τομή αυτή είναι κενή. Ακόμη, προτείνεται μια πιο ακριβής εκτίμηση της χρονικής στιγμής εμφάνισης του σφάλματος, ενώ παρατίθενται οι συνθήκες υπό τις οποίες ένα σφάλμα μπορεί να μην ανιχνευθεί ποτέ με την εφαρμογή των προτεινόμενων μεθόδων.
Η συγκεκριμένη διατριβή επίσης ερευνά την αποτελεσματικότητα της προτεινόμενης μεθοδολογίας Ανίχνευσης και Διάγνωσης Σφαλμάτων για τον εντοπισμό των τρόπων εκδήλωσης σφαλμάτων σε δύο μικροηλεκτρομηχανικά συστήματα (ΜΗΜΣ), έναν ηλεκτροστατικό μικροεπενεργητή παράλληλων πλακών και ένα ατομικό μικροσκόπιο συντονισμού στρέψης. Από πλευράς μηχανικής, οι τρόποι εκδήλωσης σφαλμάτων στα δομικά στοιχεία του μικροεπενεργητή ή του ατομικού μικροσκοποίου αντιμετωπίζονται ως απότομες παραμετρικές, οι οποίες εντοπίζονται και διαγιγνώσκονται από τις προτεινόμενες μεθόδους.
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Whiskers: The Role of Electric Fields in the Formation Mechanism and Methods for Whisker Growth MitigationBorra, Venkata Shesha Vamsi January 2017 (has links)
No description available.
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