Peanut lectin binds to a subpopulation of mitochondria-rich cells in the rainbow trout gill epithelium

2001 ◽  
Vol 281 (5) ◽  
pp. R1718-R1725 ◽  
Author(s):  
Greg G. Goss ◽  
Sophia Adamia ◽  
Fernando Galvez
Keyword(s):  
Electron Microscopy ◽  
Rainbow Trout ◽  
Chloride Cells ◽  
Isolated Cells ◽  
Peanut Lectin ◽  
Pavement Cells ◽  
Transmission Electron ◽  
Percoll Density Gradient ◽  
Trout Gill

Fluorescently labeled peanut lectin agglutinin (PNA-FITC) was used to identify a subtype of mitochondria-rich (MR) cells in the gills of freshwater rainbow trout. In situ binding of PNA-FITC was visualized by inverted fluorescence microscopy and found to bind to cells on the trailing edge of the filament epithelium as demonstrated by differential interference contrast optics. The amount of PNA-FITC binding on the filament epithelium increased with cortisol pretreatment concomitant with an increased chloride cell fractional area as demonstrated by scanning electron microscopy. Dispersed gill cells were isolated by trypsinization and separated using a discontinuous Percoll density gradient. Cells migrating to the 1.06–1.09 g/ml interface were found to be MR as demonstrated by staining with the vital mitochondrial dye 4-(4-(dimethylamino)styryl)- N-methylpyridinium iodide and transmission electron microscopy (TEM). However, only ∼40% of the MR cells were found to bind PNA-FITC. Cortisol pretreatment increased the relative numbers of MR cells isolated from the dispersed gill cell population, but the relative proportions of PNA binding cells remained unchanged. Ultrastructural analysis of isolated cells in the TEM demonstrated that the MR cell fraction was comprised of a mixed population of chloride cells and pavement cells.

scholarly journals Immunolocalization of proton pumps (H+-ATPase) in pavement cells of rainbow trout gill

1995 ◽  
Vol 198 (12) ◽  
pp. 2619-2629 ◽  
Author(s):  
G Sullivan ◽  
J Fryer ◽  
S Perry
Keyword(s):  
Electron Microscopy ◽  
Rainbow Trout ◽  
Epithelial Cells ◽  
Apical Membrane ◽  
Respiratory Acidosis ◽  
Proton Pumps ◽  
Ion Regulation ◽  
Western Blots ◽  
Pavement Cells ◽  
Trout Gill

The expression of the V-type proton ATPase (H+-ATPase) was examined in the gill of the freshwater rainbow trout (Oncorhynchus mykiss) using immunocytochemistry in concert with laser scanning confocal or electron microscopy. A synthetic peptide consisting of the carboxy-terminal region of the 31 kDa subunit of the bovine renal H+-ATPase was used to generate an antiserum in rabbits, and its suitability for use in trout gill was confirmed by western blotting. Gill epithelial cells demonstrated specific immunoreactivity, the intensity of which was increased markedly after 18 h of exposure to hypercapnia (1 % CO2 in air). The increased intensity of H+-ATPase immunoreactivity was associated with elevated branchial net acid excretion. In the hypercapnic fish, the specific immunoreactivity was associated with both the apical membrane and cytoplasm. Electron microscopy revealed that specific immunoreactivity was localized to the pavement cells and was particularly associated with the apical membrane and subapical cytoplasmic vesicles. The increased H+-ATPase immunoreactivity in the epithelial cells of hypercapnic fish and the increased intensity of the immunoreactive bands in western blots from hypercapnic fish demonstrate an 'up-regulation' of this protein in response to respiratory acidosis. The results are discussed with reference to current models of acid­base and ion regulation in the gill of freshwater fish.

Isolation and characterization of mitochondria-rich cell types from the gill of freshwater rainbow trout

2002 ◽  
Vol 282 (3) ◽  
pp. R658-R668 ◽  
Author(s):  
Fernando Galvez ◽  
Scott D. Reid ◽  
Guy Hawkings ◽  
Greg G. Goss
Keyword(s):  
Rainbow Trout ◽  
Iron Particle ◽  
Cell Types ◽  
Chloride Cells ◽  
Transmission Electron Microscopy Analysis ◽  
Pavement Cells ◽  
Isolation And Characterization ◽  
Transmission Electron ◽  
Electron Microscopy Analysis ◽  
Magnetic Cell Separation

A magnetic cell separation technique (MACS) was developed for isolating and characterizing peanut lectin agglutinin positive (PNA+) cells from rainbow trout gills. Percoll density separated mitochondria-rich (MR) cells were serially labeled with PNA-FITC and an anti-FITC antibody covalently coupled to a 50-nm iron particle and then applied to a magnetic column. PNA+ MR cells were enriched to >95% purity. Transmission electron microscopy analysis of both the PNA+ and PNA negative (PNA−) fraction showed that PNA binds to MR chloride cells while the PNA− cell fraction is comprised of MR cells with features characteristic of pavement cells. Western blotting demonstrated that both PNA+ and PNA− fractions had high levels of Na+-K+-ATPase and Sco1 expression; however, relative expression of H+-ATPase in PNA+ and PNA− cells demonstrated that untreated fish had twofold higher H+-ATPase levels in PNA− cells relative to the PNA+ cells. Furthermore, hypercapnic acidosis significantly increased the relative H+-ATPase expression on PNA− cells only, whereas metabolic alkalosis had no significant effect.

In situ observations of electromigration induced void behavior

1995 ◽  
Vol 53 ◽  
pp. 244-245
Author(s):  
T. Marieb ◽  
J. C. Bravman ◽  
P. Flinn ◽  
D. Gardner ◽  
M. Madden
Keyword(s):  
Electron Microscopy ◽  
Void Nucleation ◽  
Plan View ◽  
Transmission Electron ◽  
Nucleation Sites ◽  
Microelectronics Industry ◽  
In Situ Observations ◽  
Backscatter Electron Detector ◽  
Voltage Scanning

Electromigration and stress voiding have been active areas of research in the microelectronics industry for many years. While accelerated testing of these phenomena has been performed for the last 25 years[1-2], only recently has the introduction of high voltage scanning electron microscopy (HVSEM) made possible in situ testing of realistic, passivated, full thickness samples at high resolution.With a combination of in situ HVSEM and post-testing transmission electron microscopy (TEM) , electromigration void nucleation sites in both normal polycrystalline and near-bamboo pure Al were investigated. The effect of the microstructure of the lines on the void motion was also studied.The HVSEM used was a slightly modified JEOL 1200 EX II scanning TEM with a backscatter electron detector placed above the sample[3]. To observe electromigration in situ the sample was heated and the line had current supplied to it to accelerate the voiding process. After testing lines were prepared for TEM by employing the plan-view wedge technique [6].

Epitaxy and texture analysis of Bi-Sr-Ca-Cu-O thin films on (100) MgO using Transmission Electron Microscopy

1990 ◽  
Vol 48 (4) ◽  
pp. 68-69
Author(s):  
J. T. Sizemore ◽  
D. G. Schlom ◽  
Z. J. Chen ◽  
J. N. Eckstein ◽  
I. Bozovic ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Thin Films ◽  
Transmission Electron Microscopy ◽  
Critical Currents ◽  
Unit Cell ◽  
X Ray Diffraction ◽  
Cell Axis ◽  
Transmission Electron ◽  
Synthesis Procedure

Investigators observe large critical currents for superconducting thin films deposited epitaxially on single crystal substrates. The orientation of these films is often characterized by specifying the unit cell axis that is perpendicular to the substrate. This omits specifying the orientation of the other unit cell axes and grain boundary angles between grains of the thin film. Misorientation between grains of YBa2Cu3O7−δ decreases the critical current, even in those films that are c axis oriented. We presume that these results are similar for bismuth based superconductors and report the epitaxial orientations and textures observed in such films.Thin films of nominally Bi2Sr2CaCu2Ox were deposited on MgO using molecular beam epitaxy (MBE). These films were in situ grown (during growth oxygen was incorporated and the films were not oxygen post-annealed) and shuttering was used to encourage c axis growth. Other papers report the details of the synthesis procedure. The films were characterized using x-ray diffraction (XRD) and transmission electron microscopy (TEM).

Remote On-Line Control of a High-Voltage in situ Transmission Electron Microscope with A Rational User Interface

1996 ◽  
Vol 54 ◽  
pp. 384-385
Author(s):  
M.A. O’Keefe ◽  
J. Taylor ◽  
D. Owen ◽  
B. Crowley ◽  
K.H. Westmacott ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
User Interface ◽  
Transmission Electron Microscope ◽  
High Voltage ◽  
Materials Science ◽  
Transmission Electron ◽  
On Line ◽  
Electron Microscopes

Remote on-line electron microscopy is rapidly becoming more available as improvements continue to be developed in the software and hardware of interfaces and networks. Scanning electron microscopes have been driven remotely across both wide and local area networks. Initial implementations with transmission electron microscopes have targeted unique facilities like an advanced analytical electron microscope, a biological 3-D IVEM and a HVEM capable of in situ materials science applications. As implementations of on-line transmission electron microscopy become more widespread, it is essential that suitable standards be developed and followed. Two such standards have been proposed for a high-level protocol language for on-line access, and we have proposed a rational graphical user interface. The user interface we present here is based on experience gained with a full-function materials science application providing users of the National Center for Electron Microscopy with remote on-line access to a 1.5MeV Kratos EM-1500 in situ high-voltage transmission electron microscope via existing wide area networks. We have developed and implemented, and are continuing to refine, a set of tools, protocols, and interfaces to run the Kratos EM-1500 on-line for collaborative research. Computer tools for capturing and manipulating real-time video signals are integrated into a standardized user interface that may be used for remote access to any transmission electron microscope equipped with a suitable control computer.

Transmission Electron Microscopy of Epitaxial silicides grown by ultra high vacuum deposition

1989 ◽  
Vol 47 ◽  
pp. 462-463
Author(s):  
D. Loretto ◽  
J. M. Gibson ◽  
S. M. Yalisove
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Electron Diffraction ◽  
High Vacuum ◽  
High Energy ◽  
Energy Electron ◽  
Ultra High Vacuum ◽  
Ex Situ ◽  
Transmission Electron

The silicides CoSi2 and NiSi2 are both metallic with the fee flourite structure and lattice constants which are close to silicon (1.2% and 0.6% smaller at room temperature respectively) Consequently epitaxial cobalt and nickel disilicide can be grown on silicon. If these layers are formed by ultra high vacuum (UHV) deposition (also known as molecular beam epitaxy or MBE) their thickness can be controlled to within a few monolayers. Such ultrathin metal/silicon systems have many potential applications: for example electronic devices based on ballistic transport. They also provide a model system to study the properties of heterointerfaces. In this work we will discuss results obtained using in situ and ex situ transmission electron microscopy (TEM).In situ TEM is suited to the study of MBE growth for several reasons. It offers high spatial resolution and the ability to penetrate many monolayers of material. This is in contrast to the techniques which are usually employed for in situ measurements in MBE, for example low energy electron diffraction (LEED) and reflection high energy electron diffraction (RHEED), which are both sensitive to only a few monolayers at the surface.

Devitrification products of rapidly solidified Ni-Nb amorphous alloys.

1990 ◽  
Vol 48 (4) ◽  
pp. 950-951
Author(s):  
G. A. Bertero ◽  
W.H. Hofmeister ◽  
N.D. Evans ◽  
J.E. Wittig ◽  
R.J. Bayuzick
Keyword(s):  
Electron Microscopy ◽  
Amorphous Structure ◽  
Sulphuric Acid Solution ◽  
X Ray Diffraction ◽  
High Fracture ◽  
Transmission Electron ◽  
Xrd Techniques ◽  
Rapidly Solidified ◽  
Electromagnetically Levitated

Rapid solidification of Ni-Nb alloys promotes the formation of amorphous structure. Preliminary results indicate promising elastic properties and high fracture strength for the metallic glass. Knowledge of the thermal stability of the amorphus alloy and the changes in properties with temperature is therefore of prime importance. In this work rapidly solidified Ni-Nb alloys were analyzed with transmission electron microscopy (TEM) during in-situ heating experiments and after isothermal annealing of bulk samples. Differential thermal analysis (DTA), scanning electron microscopy (SEM) and x-ray diffraction (XRD) techniques were also used to characterize both the solidification and devitrification sequences.Samples of Ni-44 at.% Nb were electromagnetically levitated, melted, and rapidly solidified by splatquenching between two copper chill plates. The resulting samples were 100 to 200 μm thick discs of 2 to 3 cm diameter. TEM specimens were either ion-milled or alternatively electropolished in a methanol-10% sulphuric acid solution at 20 V and −40°C.

Investigation of Switching Mechanism in HfO2-Based Oxide Resistive Memories by In-Situ Transmission Electron Microscopy and Electron Energy Loss Spectroscopy

2017 ◽  
Author(s):  
T. Dewolf ◽  
D. Cooper ◽  
N. Bernier ◽  
V. Delaye ◽  
A. Grenier ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Energy Loss ◽  
Electron Energy ◽  
Electron Energy Loss Spectroscopy ◽  
Hafnium Dioxide ◽  
Switching Mechanism ◽  
Transmission Electron ◽  
Electron Energy Loss

Abstract Forming and breaking a nanometer-sized conductive area are commonly accepted as the physical phenomenon involved in the switching mechanism of oxide resistive random access memories (OxRRAM). This study investigates a state-of-the-art OxRRAM device by in-situ transmission electron microscopy (TEM). Combining high spatial resolution obtained with a very small probe scanned over the area of interest of the sample and chemical analyses with electron energy loss spectroscopy, the local chemical state of the device can be compared before and after applying an electrical bias. This in-situ approach allows simultaneous TEM observation and memory cell operation. After the in-situ forming, a filamentary migration of titanium within the dielectric hafnium dioxide layer has been evidenced. This migration may be at the origin of the conductive path responsible for the low and high resistive states of the memory.

scholarly journals Fabrication of a liquid cell for in situ transmission electron microscopy

Microscopy ◽  
2020 ◽  
Author(s):  
Xiaoguang Li ◽  
Kazutaka Mitsuishi ◽  
Masaki Takeguchi
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Cost Effective ◽  
Production Method ◽  
Microscopy Imaging ◽  
Transmission Electron ◽  
In Situ Electron Microscopy ◽  
Liquid Cell ◽  
Si Wafer

Abstract Liquid cell transmission electron microscopy (LCTEM) enables imaging of dynamic processes in liquid with high spatial and temporal resolution. The widely used liquid cell (LC) consists of two stacking microchips with a thin wet sample sandwiched between them. The vertically overlapped electron-transparent membrane windows on the microchips provide passage for the electron beam. However, microchips with imprecise dimensions usually cause poor alignment of the windows and difficulty in acquiring high-quality images. In this study, we developed a new and efficient microchip fabrication process for LCTEM with a large viewing area (180 µm × 40 µm) and evaluated the resultant LC. The new positioning reference marks on the surface of the Si wafer dramatically improve the precision of dicing the wafer, making it possible to accurately align the windows on two stacking microchips. The precise alignment led to a liquid thickness of 125.6 nm close to the edge of the viewing area. The performance of our LC was demonstrated by in situ transmission electron microscopy imaging of the dynamic motions of 2-nm Pt particles. This versatile and cost-effective microchip production method can be used to fabricate other types of microchips for in situ electron microscopy.

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