Microwave Processing of Cell Monolayers in Situ for Postembedding Immunocytochemistry with Retention of Ultrastructure and Antigenicity

1998 ◽  
Vol 4 (S2) ◽  
pp. 854-855
Author(s):  
Victoria J. Madden

Increasingly, cells isolated from blood and body fluids and cells grown in culture are becoming the experimental models of choice for biological research. The demand for demonstrating biochemical processes morphologically is also becoming commonplace in the electron microscopy laboratory. Successful fixation, in situ embedment, and ultrathinsectioning of cell monolayers can be difficult to achieve for routine transmission electron microscopy. For postembedding immunocytochemistry, processing becomes more complex due to fixation constraints and the use of acrylic resins. The object of this paper is to present a reliable, rapid method for processing monolayers that preserves both the ultrastructure of the cells and antigenicity.The equipment used for this procedure was a Pelco Model 3440 MAX laboratory microwave oven equipped with a temperature probe and a maximum power output of 800 watts. Using a neon bulb array, the oven cavity is calibrated to determine the microwave energy distribution.

Author(s):  
T. Marieb ◽  
J. C. Bravman ◽  
P. Flinn ◽  
D. Gardner ◽  
M. Madden

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].


Author(s):  
J. T. Sizemore ◽  
D. G. Schlom ◽  
Z. J. Chen ◽  
J. N. Eckstein ◽  
I. Bozovic ◽  
...  

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).


Author(s):  
M.A. O’Keefe ◽  
J. Taylor ◽  
D. Owen ◽  
B. Crowley ◽  
K.H. Westmacott ◽  
...  

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.


Author(s):  
D. Loretto ◽  
J. M. Gibson ◽  
S. M. Yalisove

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.


Author(s):  
G. A. Bertero ◽  
W.H. Hofmeister ◽  
N.D. Evans ◽  
J.E. Wittig ◽  
R.J. Bayuzick

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.


Author(s):  
T. Dewolf ◽  
D. Cooper ◽  
N. Bernier ◽  
V. Delaye ◽  
A. Grenier ◽  
...  

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.


Microscopy ◽  
2020 ◽  
Author(s):  
Xiaoguang Li ◽  
Kazutaka Mitsuishi ◽  
Masaki Takeguchi

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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