scholarly journals Platelet Activation State Intermixing in a Venous Puncture Model Indicates Novel Patterns of Thrombus Formation

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 9-9
Author(s):  
Brian Storrie ◽  
Sung W. Rhee ◽  
Irina D Pokrovskaya ◽  
Kenny Ling ◽  
Yajnesh Vedanaparti ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Platelet Activation ◽  
Vessel Wall ◽  
Jugular Vein ◽  
Thrombus Formation ◽  
Sem Images ◽  
Puncture Wound ◽  
Activated Platelets ◽  
Puncture Model ◽  
Scanning Electron

Introduction: Platelet recruitment to generate a thrombus is key to bleeding cessation. That recruitment is dependent on a series of platelet activation processes that include adhesion to the exposed vessel matrix, platelet-platelet adhesion and platelet granule release. How platelet activation is patterned to generate a thrombus has previously been studied by intravital light microscopy, two-photon microscopy and scanning electron microscopy at resolutions insufficient to infer platelet activation at the level of the individual platelet. Here, we present a collaborative effort to stratify spatially the extent of platelet activation at the cellular level in a mouse jugular vein puncture model. We used wide-area transmission electron microscopy (WA-TEM) and serial block face scanning electron microscopy (SBF-SEM) at a resolution of 3 to 100 nm across whole thrombi to determine activation state of individual platelets. Our results, indicate a pattern of platelet stratification within the puncture wound that varies in a time-dependent manner with distinct structural stages in the formation of the thrombus. Methods: Jugular vein thrombi from C57BL/6 mice were collected 1, 5, or 20 min after a 300 µm needle puncture and prepared for EM imaging. For WA-TEM, hundreds of overlapping 3 nm resolution images were acquired using a gondimeter stage. The images sets were aligned using NIH Fiji software to create a single high-res, thrombus-wide image. Individual platelets were stratified into morphologically defined activation states (1: no activation, 2: decreased granule number, 3: no visible granules left, 4: hollow inside). The spatial distribution of platelet stratification was analyzed using iVision software. For SBF-SEM, 100-nm XY-resolution SEM images were collected every 200 nm and 20 nm XY-resolution images every 20 µm. Semi-automated stratification of platelet activation state in individual slices of the thrombus were combined into a 3-D representation using Amira software. Volumetric distributions of platelets with respect to the puncture hole and the vascular wall were quantified. Results: Thrombus Formation Stage 1 (anchor and extend) -- One min post puncture, platelets were anchored in clumps along the exposed vessel wall. Near the damaged vessel wall was a peripheral layer of activated or degranulated platelets (states 3 and 4) covered by additional layers of less-activated platelets (state 1 and 2). Short cylindrical ingrowths extended into the 300 µm hole. Unexpectedly, large aggregates of platelets with a mixture of activation states (states 1 - 4) were found extending from these anchor points into the hole and vertically into the intravascular space. Aggregate surface layers were composed mostly of degranulated platelets (states 3 and 4). Less than 40% of neighboring platelets were of the same activation state as their neighbor. Surprisingly, <2% of platelet-occupied volume within the puncture hole contained largely degranulated platelets (aggregates of only states 3 and 4). Stage 2 (cap and erect) -- At 5 min after injury, the puncture hole was capped. ~70% of platelets neighboring degranulated platelets (3 and 4) formed visible aggregates within the thrombus. These aggregates were found along the exposed vessel wall and encasing vertical platelet aggregate towers containing a mixture of platelets in different states (1 - 4). Towers were typically separated by large cavities. SBF-SEM images, a machine-based, unbiased sampling of the underlying platelet distribution, revealed that ~10% of the platelet volume in the puncture hole of the thrombus and the intravascular towers contained largely degranulated platelets, similar to the data from WA-TEM. Stage 3 (infill and remodeling) - At 20 min post-puncture, the thrombus was filled with a mixture of platelets of varying activation states, which surrounded central, vertical aggregates (towers) of degranulated platelets seen at 5 min. Only minor cavity space was apparent. The intravascular surface of the thrombus was covered with an ~10 platelet-thick layer of loosely packed, variably activated platelets (states 1 - 4). Conclusions: Our results demonstrate dynamic spatial patterns of platelet activation within a forming puncture-wound thrombus. Such patterns yield insights into thrombus formation and suggest the need to reference platelets defects and anti-thrombotics drugs against new models. Figure Disclosures No relevant conflicts of interest to declare.

scholarly journals Platelet Aggregation and Fibrin Deposition: The Yin and Yang of Thrombin Activity Regulation By Platelet Glycoprotein Ibα

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 114-114
Author(s):  
Alessandro Zarpellon ◽  
Patrizia Marchese ◽  
Antonella Zampolli ◽  
Grazia Loredana Mendolicchio ◽  
Zaverio M. Ruggeri
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Shear Stress ◽  
Carotid Artery ◽  
Platelet Activation ◽  
Ex Vivo ◽  
Thrombus Formation ◽  
Platelet Glycoprotein ◽  
Type I ◽  
Scanning Electron

Abstract Generation of α-thrombin (FIIa) in response to vascular injury is a key mechanism influencing thrombus formation. Platelet activation by FIIa is mediated by different protease activated receptors (PARs), although the most abundant FIIa binding site on platelets, but not a substrate for proteolysis, is glycoprotein (GP) Ibα in the GPIb-IX-V complex. The functional role of GPIbα in mediating/regulation thrombin functions relative to that of different PARs remains unclear. The goal of these studies was to define how binding to GPIbα can modulate FIIa functions. In mouse platelets we replaced endogenous GPIbα with either its human wild type counterpart (huGPIbα-WT) or with huGPIbα mutated at key residues involved in thrombin binding (D277N, Y276-8-9/F). Because these two mutations resulted in an undistinguishable phenotype, they are designated collectively as huGPIbα-Mut hereon. Mice expressing huGPIbα, WT or Mut, were evaluated in intravital models of arterial thrombosis induced by a ferric chloride-induced carotid artery lesion and venous thromboembolism induced by intravenous α-thrombin injection. Moreover, the blood of huGPIbα WT or Mut mice was also tested in an ex vivo model of thrombus formation upon perfusion over a thrombogenic surface under controlled flow conditions and platelets were evaluated for their responses to FIIa-induced activation. Mice expressing huGPIbα - WT or Mut - have comparable platelet counts and GPIbα surface density. Moreover, huGPIbα-WT platelets bind FIIa similarly than their normal human control counterpart, while huGPIbα-Mut platelets have essentially no detectable FIIa binding. Upon FIIa stimulation, which on mouse platelets is mediated by PAR4, aggregation and Ca2+ transients were significantly enhanced in huGPIbα-Mut as compared to huGPIbα-WT. In contrast, blocking FIIa binding to GPIbα on human platelets essentially abolished FIIa mediated activation, which in human occurs predominantly through PAR1. These results are compatible with the conclusion that, in mice, GPIbα is a competitive inhibitor of FIIa for PAR4-mediated functions. In the presence of metabolically inactive (PGE1 treated) huGPIbα-Mut washed platelets, the clotting time of a purified fibrinogen solution was significantly shorter when triggered by relative high concentration of FIIa (4 nM), but pronouncedly prolonged at a lower FIIa concentration (0.5 nM). Clot visualization showed a much more structured fibrin mesh in the presence of huGPIbα-WT platelets, which was lost with in the presence of huGPIbα-Mut platelets. Mutant mice tested in a model of carotid artery injury exhibited a pronounced prothrombotic phenotype, with a shorter time to occlusion. However they were protected from death induced by I.V. injection of α-thrombin. In ex vivo perfusion studies, the total volume of platelet aggregates formed in huGPIbα-Mut mouse blood exposed to acid-insoluble fibrillar collagen type I was slightly bigger than in huGPIbα-WT mice, but the number of thrombi was increased and their individual size smaller. These huGPIbα-Mut platelets exhibited clear signs of increased activation, as visualized by scanning electron microscopy (SEM). Strikingly, fibrin was almost totally absent in the huGPIbα-Mut thrombi. This was in striking contrast with what observed in huGPIbα-WT mice, in which the surface of platelet thrombi with directly and tightly connected with thick fibrin fibers as visualized by scanning electron microscopy. Possibly because of the reduced platelet membrane-fibrin fibril connection in huGPIbα-Mut platelets, these mice were significantly less susceptible to death when injected with an α-thrombin dose that caused 80% mortality plus in huGPIbα-WT mice. Thus, mice whose platelets have defective α-thrombin binding to GPIbα have a prothrombotic phenotype in high shear stress flow arteries and are protected from thromboembolic death in the low shear stress venous circulation. Our findings identify GPIbα as a relevant FIIa activity modulator in hemostasis and thrombosis through distinct and opposite mechanisms affecting platelet activation (The Yin) and fibrin formation (The Yang). Disclosures No relevant conflicts of interest to declare.

scholarly journals Bleeding Cessation in a Mouse Jugular Vein Puncture Wound Model Is Caused By Extravascular Capping, Not Hole Infill

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 13-14
Author(s):  
Brian Storrie ◽  
Sung Rhee ◽  
Timothy J. Stalker ◽  
Irina D Pokrovskaya ◽  
Kenny Ling ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Shell Model ◽  
Mouse Models ◽  
Thrombus Formation ◽  
Vascular Damage ◽  
Puncture Wound ◽  
Platelet Aggregates ◽  
Granule Secretion ◽  
Scanning Electron

Introduction: Vascular damage comes in many forms with the puncture wound likely the longest known to humans. Experimentally, vascular damage has typically been visualized in mouse models in which there is little bleeding. Under these conditions, damage is limited mostly to the endothelial layer lining the vessel. Visualization has varied from light to scanning electron microscopy. Interpretation has been dominated by intravital microscopy outcomes in which an initial layer of p-selection-exposed, i.e., α-granule secretion-positive, platelets is deposited in association with the damaged vessel wall and extended by the accumulation of a less activated outer layer of platelets. These results have given rise to a Core and Shell model of platelet-rich thrombus formation [Tomaiuolo et al., 2017]. Recently, new mouse models have been presented in which the vessel is punctured to create a 300 to 600 micron wound hole [Tomoiuolo et al., 2019]. Bleeding is now profuse. The puncture wound results have been interpreted within a Core and Shell model. However, two important aspects of the experimental data [Tomoiuolo et al., 2019] suggest that the existing model may not explain the actual results. First, p-selectin expression as a marker for α-granule secretion and platelet activation was present in limited areas towards the periphery of the resulting thrombus, not as well-defined Core. Second, the hemostatic thrombus when viewed at early stages, ex vivo, showed a pebbly distribution of platelet aggregates suggestive of nucleated platelet accumulation rather than the smooth layers that would follow from a Core and Shell model. We hypothesize that nucleated accumulation of platelet aggregates within the puncture hole could provide pedestals upon which localized accumulation of platelets form the infrastructure of a vaulted thrombus whose extravascular capping leads to bleeding cessation. Methods: To test the proposed hypothesis, we visualized the interior and overall structure of the forming puncture wound thrombus in full 3D at sub-platelet level resolution. To achieve this end, we took our proven serial block face scanning electron microscopy (SBF-SEM) protocols for visualizing platelet organelles in 3D [Pokrovskaya et al., 2018] and adapted them to the visualization of forming thrombi over 1000s of image. To localize samples for electron microscopy, we used in vivo antibody labeling [Tomaiuolo et al., 2019]. This approach had the added advantage of enabling correlative light microscopy mapping overall p-selectin, a marker of platelet secretion, and fibrin distributions against 3D, platelet resolution, thrombus morphology. Results: We found that a 1 min thrombus, pre-bleeding cessation, was structured about the localized accumulation of pedestal-like platelet aggregates along the sides of the puncture hole, extended and spaced along the extravascular adventitia. Subsequent pedestal extension formed a "pontoon" bridge that "capped" extravascularly the puncture hole. At 5 min, full bleeding cessation, we found that forming platelet thrombus had a Swiss cheese-like interior of vaults that were continuous with the intravascular vessel lumen and framed by columns of platelets, presumed pedestal extensions. The thrombus was sealed on the extravascular side by a platelet "cap" (Figure). As expected after bleeding cessation, red blood cells accumulated on the intravascular side of the cap. Formation of a tightly sealed cap was dependent on α-granule secretion as indicated by the effect of knockout of VAMP-8, the primary SNARE protein involved in a-granule release. Based upon morphology, vaults within the forming thrombus were lined with apparent procoagulant platelets providing a potential protected surface for coagulation factor activation. Conclusions: We conclude that bleeding cessation in a true puncture wound occurs from the extravascular side of the thrombus rather than through the formation of a platelet plug that fills the hole. We propose an alternative model of bleeding cessation in which localized platelet aggregates are the starting pedestal upon which all subsequent steps in puncture thrombus formation build, i.e., "Cap and Build". The extent to which properties differ among systems remains an open question. Tomaiuolo et al. 2017. Intervent. Cardiol. Clin. 6: 1-12. Pokrovskaya et al. 2018. Blood Adv. 2: 2947-2958 Tomaiuolo et al. 2019. Proc. Natl. Acad. Sci. USA 116:2 243-2252 Figure Disclosures No relevant conflicts of interest to declare.

The Effect of Interleukin-1 on Venous Endothelium – An Ultrastructural Study

1991 ◽  
Vol 66 (06) ◽  
pp. 725-729 ◽  
Author(s):  
R E Merton ◽  
D Hockley ◽  
E Gray ◽  
S Poole ◽  
D P Thomas
Keyword(s):  
Electron Microscopy ◽  
Jugular Vein ◽  
Interleukin 1 ◽  
The Other ◽  
Cell Junctions ◽  
Fibrin Deposition ◽  
Activated Platelets ◽  
Vein Segment ◽  
Isolated Segment ◽  
Scanning Electron

SummaryThe effect of systemic interleukin-1 (IL-1) on venous endothelium in the presence and absence of stasis has been studied by scanning electron microscopy (SEM). Recombinant human IL-1β at a concentration of 1 εg/kg or saline was injected intravenously into rabbits and allowed to circulate for 0.5 or 4.0 h after which complete stasis was induced for 1 h in an isolated segment of each jugular vein. One vein segment was then excised and the contents examined macroscopically for thrombi, while the other segment was fixed for SEM examination. When examined by SEM the endothelium from rabbits injected with IL-1β was perturbed with increased surface microvilli, blebs and gaps at cell junctions when compared with saline controls. Fibrin deposition was also observed after IL-1β, as was the adherence of essentially non-activated platelets to intact endothelium. However, macroscopic thrombi were not formed in isolated vein segments. We conclude that although fibrin strands and platelets were deposited on the endothelium, IL-1 is not a sufficiently powerful procoagulant stimulus to lead to an occlusive thrombus in acute experiments.

Recruitment of Labelled Monocytes by Experimental Venous Thrombi

2001 ◽  
Vol 85 (06) ◽  
pp. 1018-1024 ◽  
Author(s):  
C. L. McGuinness ◽  
J. Humphries ◽  
M. Waltham ◽  
K. G. Burnand ◽  
M. Collins ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Vessel Wall ◽  
Time Course ◽  
Thrombus Formation ◽  
Vena Cava ◽  
Steady Increase ◽  
Time Interval ◽  
Venous Thrombus ◽  
Scanning Electron

Summary Objective. Infusing monocytes that have been stimulated to produce fibrinolytic activators and factors that regulate cell proliferation, migration and maturation, might enhance venous thrombus resolution. The aim of this study was to determine the time course of infused monocyte recruitment into venous thrombus in an appropriate model of this disease. Design and Methods. Thrombus was induced in the inferior vena cava of male Wistar rats using reduced flow (80-90% stenosis). The vessel wall was examined at 1hr by scanning electron microscopy. Resolving thrombi with surrounding vena cava were obtained at 1, 7, 14 and 21 days after induction (n = 8). Sections, taken at 0.5 mm intervals (10-15 sections per thrombus), were stained using haematoxylin, Martius Scarlet Blue and antibodies against monocytes, platelets and fibrin. Sections from human venous thrombi (n = 4) were similarly stained. The area occupied by monocytes (in relative pixel units, RPU) was determined using computer aided image analysis. Peripheral rat blood monocytes were extracted, fluorescently labelled and injected intravenously into 7 rats prior to thrombus induction. Vena cava with thrombus was harvested 1 h, 2, 3, 4, 7, 14 and 25 days after induction and their fluorescence measured. The fluorescent content of the caval wall and thrombus was analysed in greater detail at 2 and 25 days after thrombus induction (n = 4 at each time interval). Results. Experimental thrombi were structurally similar to human thrombus and resolved within 14-21 days. Scanning electron microscopy showed minimal endothelial damage at 1 h with signs of early thrombus formation (platelet, red cell leukocyte and fibrin deposition). Neutrophils were the predominant leukocyte in the thrombus at 1 day, with monocytes making up only 0.3% (0.04% sem) of the area of the thrombus. There was a steady increase in thrombus monocyte content and by 21 days the percentage area of thrombus covered by monocytes had increased by over 35 fold to 11.5% (2.3% sem) (p <0.001). Initially, monocytes appeared around the edge of the thrombus and became more evenly distributed through the thrombus as resolution progressed. Labelled monocytes could be found in the circulation up to 1 week after infusion. The fluorescent content (RPU) of the thrombus increased over 25 days (mean RPU At 2 days 0.012, sem 0.005; mean RPU at 25 days 1.062, sem 0.252, p = 0.008). The number of labelled monocytes in the vessel wall peaked at 2 days and decreased thereafter.

Neutrophil Derived Cathepsin G Induces Potentially Thrombogenic Changes in Human Endothelial Cells: a Scanning Electron Microscopy Study in Static and Dynamic Conditions

1994 ◽  
Vol 72 (01) ◽  
pp. 140-145 ◽  
Author(s):  
Valeri Kolpakov ◽  
Maria Cristina D'Adamo ◽  
Lorena Salvatore ◽  
Concetta Amore ◽  
Alexander Mironov ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Thrombus Formation ◽  
Cathepsin G ◽  
Arterial Segment ◽  
Dynamic Conditions ◽  
Activated Neutrophils ◽  
Scanning Electron

SummaryActivated neutrophils may promote thrombus formation by releasing proteases which may activate platelets, impair the fibrinolytic balance and injure the endothelial monolayer.We have investigated the morphological correlates of damage induced by activated neutrophils on the vascular wall, in particular the vascular injury induced by released cathepsin G in both static and dynamic conditions.Human umbilical vein endothelial cells were studied both in a cell culture system and in a model of perfused umbilical veins. At scanning electron microscopy, progressive alterations of the cell monolayer resulted in cell contraction, disruption of the intercellular contacts, formation of gaps and cell detachment.Contraction was associated with shape change of the endothelial cells, that appeared star-like, while the underlying extracellular matrix, a potentially thrombogenic surface, was exposed. Comparable cellular response was observed in an “in vivo” model of perfused rat arterial segment. Interestingly, cathepsin G was active at lower concentrations in perfused vessels than in culture systems. Restoration of blood flow in the arterial segment previously damaged by cathepsin G caused adhesion and spreading of platelets on the surface of the exposed extracellular matrix. The subsequent deposition of a fibrin network among adherent platelets, could be at least partially ascribed to the inhibition by cathepsin G of the vascular fibrinolytic potential.This study supports the suggestion that the release of cathepsin G by activated neutrophils, f.i. during inflammation, may contribute to thrombus formation by inducing extensive vascular damage.

scholarly journals Microscopy Methods for Biofilm Imaging: Focus on SEM and VP-SEM Pros and Cons

Biology ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 51
Author(s):  
Michela Relucenti ◽  
Giuseppe Familiari ◽  
Orlando Donfrancesco ◽  
Maurizio Taurino ◽  
Xiaobo Li ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Ion Beam ◽  
Ruthenium Red ◽  
Variable Pressure ◽  
Sem Images ◽  
Advantages And Disadvantages ◽  
Pros And Cons ◽  
Microscopy Techniques ◽  
Scanning Electron

Several imaging methodologies have been used in biofilm studies, contributing to deepening the knowledge on their structure. This review illustrates the most widely used microscopy techniques in biofilm investigations, focusing on traditional and innovative scanning electron microscopy techniques such as scanning electron microscopy (SEM), variable pressure SEM (VP-SEM), environmental SEM (ESEM), and the more recent ambiental SEM (ASEM), ending with the cutting edge Cryo-SEM and focused ion beam SEM (FIB SEM), highlighting the pros and cons of several methods with particular emphasis on conventional SEM and VP-SEM. As each technique has its own advantages and disadvantages, the choice of the most appropriate method must be done carefully, based on the specific aim of the study. The evaluation of the drug effects on biofilm requires imaging methods that show the most detailed ultrastructural features of the biofilm. In this kind of research, the use of scanning electron microscopy with customized protocols such as osmium tetroxide (OsO4), ruthenium red (RR), tannic acid (TA) staining, and ionic liquid (IL) treatment is unrivalled for its image quality, magnification, resolution, minimal sample loss, and actual sample structure preservation. The combined use of innovative SEM protocols and 3-D image analysis software will allow for quantitative data from SEM images to be extracted; in this way, data from images of samples that have undergone different antibiofilm treatments can be compared.

scholarly journals Apolar Bioactive Fraction ofMelipona scutellarisGeopropolis onStreptococcus mutansBiofilm

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Marcos Guilherme da Cunha ◽  
Marcelo Franchin ◽  
Lívia Câmara de Carvalho Galvão ◽  
Bruno Bueno-Silva ◽  
Masaharu Ikegaki ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Biofilm Formation ◽  
Microbial Population ◽  
Ethanolic Extract ◽  
Vehicle Control ◽  
Sem Images ◽  
Killing Assay ◽  
Bioactive Fraction ◽  
Biofilm Matrix ◽  
Scanning Electron

The aim of this study was to evaluate the influence of the bioactive nonpolar fraction of geopropolis onStreptococcus mutansbiofilm. The ethanolic extract ofMelipona scutellarisgeopropolis was subjected to a liquid-liquid partition, thus obtaining the bioactive hexane fraction (HF) possessing antimicrobial activity. The effects of HF onS. mutansUA159 biofilms generated on saliva-coated hydroxyapatite discs were analyzed by inhibition of formation, killing assay, and glycolytic pH-drop assays. Furthermore, biofilms treated with vehicle control and HF were analyzed by scanning electron microscopy (SEM). HF at 250 μg/mL and 400 μg/mL caused 38% and 53% reduction in the biomass of biofilm, respectively, when compared to vehicle control (P<0.05) subsequently observed at SEM images, and this reduction was noticed in the amounts of extracellular alkali-soluble glucans, intracellular iodophilic polysaccharides, and proteins. In addition, theS. mutansviability (killing assay) and acid production by glycolytic pH drop were not affected (P>0.05). In conclusion, the bioactive HF of geopropolis was promising to control theS. mutansbiofilm formation, without affecting the microbial population but interfering with its structure by reducing the biochemical content of biofilm matrix.

scholarly journals Analysis of the type material of Gomphosphenia tackei (Bacillariophyceae) and comparison to epizoic diatom populations on freshwater snails

2021 ◽  
Vol 154 (2) ◽  
pp. 257-263
Author(s):  
Mateusz Rybak ◽  
Łukasz Peszek ◽  
Anita Poradowska
Keyword(s):  
Electron Microscopy ◽  
Lymnaea Stagnalis ◽  
Morphological Characteristics ◽  
Small Cell ◽  
Type Material ◽  
Freshwater Snails ◽  
Sem Images ◽  
Small Cell Size ◽  
Type Population ◽  
Scanning Electron

Background and aims – Hustedt (1942) originally described Gomphosphenia tackei from Germany under the name Gomphonema tackei. Because of the small cell size and the lack of scanning electron microscopy (SEM) images from the type material, it is often confused with other species from this genus, especially with G. stoermeri. The aim of this paper was to present detailed morphological characteristics of G. tackei based on the analysis of the type material and of several epizoic populations from Central Europe. Material and methods – The material in this study was collected from the shells of the freshwater snails Lymnaea stagnalis, Planorbarius corneus, and Planorbis planorbis. Additionally, for an unambiguous species identification, the type material for Gomphosphenia tackei was analyzed using light and scanning electron microscopes.Key results – The presence of Gomphosphenia tackei was confirmed in the studied material. The largest population (up to 19%) was recorded on the shell surfaces of living snails, whereas on empty shells, the diatom did not seem to be present or only in very low numbers. Valves are typically clavate with rounded apices. Valves are frequently observed in girdle view, often joint together in pairs. The valves in the studied populations had a valve length of 7–29 µm, a valve width of 3–4 µm, and a stria density of 25–29 striae in 10 µm. In the type population, valve length ranged from 7.5 to 27 µm with a valve width of 3.0–4.0 µm and a stria density of 23–29 striae per 10 µm. Striae were composed of 2–4 elongated to rounded areolae per stria. At the apices, the striae were composed of one single areola. The cells were attached to the substratum by their footpole.Conclusion – Published illustrations of Gomphosphenia tackei do not always correctly represent this species. Individual cells are attached to the substratum by secreted mucilage, probably via their areolae or girdle band pores located on the footpole.

Fabrication of Ultra Thick Ferromagnetic Structures in Silicon

2004 ◽  
Author(s):  
Debbie G. Jones ◽  
Albert P. Pisano
Keyword(s):  
Electron Microscopy ◽  
Thin Films ◽  
Scanning Electron Microscopy ◽  
Saturation Magnetization ◽  
Silicon Wafer ◽  
Fabrication Process ◽  
Sem Images ◽  
Deep Reactive Ion Etching ◽  
Scanning Electron

A novel fabrication process is presented to create ultra thick ferromagnetic structures in silicon. The structures are fabricated by electroforming NiFe into silicon templates patterned with deep reactive ion etching (DRIE). Thin films are deposited into photoresist molds for characterization of an electroplating cell. Results show that electroplated films with a saturation magnetization above 1.6 tesla and compositions of approximately 50/50 NiFe can be obtained through agitation of the electrolyte. Scanning electron microscopy (SEM) images show that NiFe structures embedded in a 500 μm thick silicon wafer are realized and the roughening of the mold sidewalls during the DRIE aids in adhesion of the NiFe to the silicon.

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