Importance of Specimen Preparation in Microscopy

2009 ◽  
pp. 289-289-19 ◽  
Author(s):  
JE Gillott
Keyword(s):  
Specimen Preparation

Direct Observation of Heat Exchanger Deposits by Cross Sectioning

1967 ◽  
Vol 25 ◽  
pp. 364-365
Author(s):  
R. W. Anderson ◽  
D. L. Senecal
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Heat Exchanger ◽  
Direct Observation ◽  
Cross Sections ◽  
Preparation Method ◽  
Specimen Preparation ◽  
Flowing Water ◽  
Transmission Electron ◽  
Deposit Layer

A problem was presented to observe the packing densities of deposits of sub-micron corrosion product particles. The deposits were 5-100 mils thick and had formed on the inside surfaces of 3/8 inch diameter Zircaloy-2 heat exchanger tubes. The particles were iron oxides deposited from flowing water and consequently were only weakly bonded. Particular care was required during handling to preserve the original formations of the deposits. The specimen preparation method described below allowed direct observation of cross sections of the deposit layers by transmission electron microscopy.The specimens were short sections of the tubes (about 3 inches long) that were carefully cut from the systems. The insides of the tube sections were first coated with a thin layer of a fluid epoxy resin by dipping. This coating served to impregnate the deposit layer as well as to protect the layer if subsequent handling were required.

Enhancement of Contrast in the CEM and STEM Using Bumpy Specimens and Employing the “Dappled Field” Mode of Operation

1974 ◽  
Vol 32 ◽  
pp. 552-553 ◽  
Author(s):  
L. Gandolfi ◽  
J. Reiffel
Keyword(s):  
Operating Mode ◽  
Scanning Transmission Electron Microscope ◽  
Dark Field ◽  
Specimen Preparation ◽  
Field Mode ◽  
Preparation Methods ◽  
Scanning Transmission Electron ◽  
Transmission Electron ◽  
Mode Of Operation ◽  
Scanning Transmission

Calculations have been performed on the contrast obtainable, using the Scanning Transmission Electron Microscope, in the observation of thick specimens. Recent research indicates a revival of an earlier interest in the observation of thin specimens with the view of comparing the attainable contrast using both types of specimens.Potential for biological applications of scanning transmission electron microscopy has led to a proliferation of the literature concerning specimen preparation methods and the controversy over “to stain or not to stain” in combination with the use of the dark field operating mode and the same choice of technique using bright field mode of operation has not yet been resolved.

Contrast from epitaxially sandwiched macromolecules

1978 ◽  
Vol 36 (2) ◽  
pp. 226-227
Author(s):  
M. Talianker ◽  
D.G. Brandon
Keyword(s):  
Gold Film ◽  
Lattice Defect ◽  
Gold Foil ◽  
Specimen Preparation ◽  
Preparation Technique ◽  
Resolution Limit ◽  
Transmission Electron ◽  
Crystal Lattice Defect ◽  
Void Effect ◽  
Lattice Resolution

A new specimen preparation technique for visualizing macromolecules by conventional transmission electron microscopy has been developed. In this technique the biopolymer-molecule is embedded in a thin monocrystalline gold foil. Such embedding can be performed in the following way: the biopolymer is deposited on an epitaxially-grown thin single-crystal gold film. The molecule is then occluded by further epitaxial growth. In such an epitaxial sandwich an occluded molecule is expected to behave as a crystal-lattice defect and give rise to contrast in the electron microscope.The resolution of the method should be limited only by the precision with which the epitaxially grown gold reflects the details of the molecular structure and, in favorable cases, can approach the lattice resolution limit.In order to estimate the strength of the contrast due to the void-effect arising from occlusion of the DNA-molecule in a gold crystal some calculations were performed.

The ionic strength dependence of chromatin folding

1978 ◽  
Vol 36 (2) ◽  
pp. 220-221
Author(s):  
F. Thoma ◽  
TH. Koller
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Ionic Strength ◽  
Specimen Preparation ◽  
Preparation Technique ◽  
Carbon Supports ◽  
Ionic Strength Dependence ◽  
Chromatin Folding ◽  
Strength Dependence ◽  
Preparation Techniques

Under a variety of electron microscope specimen preparation techniques different forms of chromatin appearance can be distinguished: beads-on-a-string, a 100 Å nucleofilament, a 250 Å fiber and a compact 300 to 500 Å fiber.Using a standardized specimen preparation technique we wanted to find out whether there is any relation between these different forms of chromatin or not. We show that with increasing ionic strength a chromatin fiber consisting of a row of nucleo- somes progressively folds up into a solenoid-like structure with a diameter of about 300 Å.For the preparation of chromatin for electron microscopy the avoidance of stretching artifacts during adsorption to the carbon supports is of utmost importance. The samples are fixed with 0.1% glutaraldehyde at 4°C for at least 12 hrs. The material was usually examined between 24 and 48 hrs after the onset of fixation.

SEM evaluation of the TEM cold-stage double-film specimen

1984 ◽  
Vol 42 ◽  
pp. 272-273
Author(s):  
Jayesh Bellare
Keyword(s):  
Spatial Distribution ◽  
Sample Preparation ◽  
Sample Thickness ◽  
Specimen Preparation ◽  
Preparation Technique ◽  
Liquid Sample ◽  
Thin Specimen ◽  
Sample Preparation Technique ◽  
Cold Stage ◽  
Film Specimen

Seeing is believing, but only after the sample preparation technique has received a systematic study and a full record is made of the treatment the sample gets.For microstructured liquids and suspensions, fast-freeze thermal fixation and cold-stage microscopy is perhaps the least artifact-laden technique. In the double-film specimen preparation technique, a layer of liquid sample is trapped between 100- and 400-mesh polymer (polyimide, PI) coated grids. Blotting against filter paper drains excess liquid and provides a thin specimen, which is fast-frozen by plunging into liquid nitrogen. This frozen sandwich (Fig. 1) is mounted in a cooling holder and viewed in TEM.Though extremely promising for visualization of liquid microstructures, this double-film technique suffers from a) ireproducibility and nonuniformity of sample thickness, b) low yield of imageable grid squares and c) nonuniform spatial distribution of particulates, which results in fewer being imaged.

Electron Microscopic Observation of Biological Specimens in Their Native State by Employing Cryogenic Techniques

1972 ◽  
Vol 30 ◽  
pp. 410-411 ◽  
Author(s):  
Tokio Nei ◽  
Haruo Yotsumoto ◽  
Yoichi Hasegawa ◽  
Yuji Nagasawa
Keyword(s):  
Electron Microscopic Observation ◽  
Surface Structures ◽  
Specimen Preparation ◽  
Native State ◽  
Electron Microscopic ◽  
Biological Specimen ◽  
Specimen Holder ◽  
Cold Stage ◽  
Preparation Techniques ◽  
Scanning Electron

In order to observe biological specimens in their native state, that is, still containing their water content, various methods of specimen preparation have been used, the principal two of which are the chamber method and the freeze method.Using its recently developed cold stage for installation in the pre-evacuation chamber of a scanning electron microscope, we have succeeded in directly observing a biological specimen in its frozen state without the need for such conventional specimen preparation techniques as drying and metallic vacuum evaporation. (Echlin, too, has reported on the observation of surface structures using the same freeze method.)In the experiment referred to herein, a small sliced specimen was place in the specimen holder. After it was rapidly frozen by freon cooled with liquid nitrogen, it was inserted into the cold stage of the specimen chamber.

On the High Voltage Electron Microscopy (1 MeV) of Biological Thick Sections

1972 ◽  
Vol 30 ◽  
pp. 464-465
Author(s):  
William H. Massover
Keyword(s):  
Electron Microscopy ◽  
High Voltage ◽  
Present Report ◽  
Biological Tissues ◽  
Specimen Preparation ◽  
Technical Problems ◽  
High Voltage Electron Microscopy ◽  
Thin Sections ◽  
Voltage Electron ◽  
High Voltage Electron

Stereoscopic examination of thick sections of fixed and embedded biological tissues by high voltage electron microscopy has been shown to allow direct visualization of three-dimensional fine structure. The present report will consider the occurrence of some new technical problems in specimen preparation and Image interpretation that are not common during lower voltage studies of thin sections.Thick Sectioning and Tissue Coloration - Epon sections of 0.5 μm or more that are cut with glass knives do not have a uniform thickness as Judged by their interference colors; these colors change with time during their flotation on the knife bath, and again when drying onto the specimen support. Quoted thicknesses thus must be considered only as rough estimates unless measured in specific regions by other methods. Chloroform vapors do not always result in good spreading of thick sections; however, they will spread spontaneously to large degrees after resting on the flotation bath for several minutes. Ribbons of thick sections have been almost impossible to obtain.

The Current Situation in Chemical Stabilization of Biological Materials

1972 ◽  
Vol 30 ◽  
pp. 132-133
Author(s):  
John H. Luft
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Osmium Tetroxide ◽  
Molecular Level ◽  
Cross Linking ◽  
Chemical Stabilization ◽  
Specimen Preparation ◽  
Sufficient Condition ◽  
Radio Telescopes

With information processing devices such as radio telescopes, microscopes or hi-fi systems, the quality of the output often is limited by distortion or noise introduced at the input stage of the device. This analogy can be extended usefully to specimen preparation for the electron microscope; fixation, which initiates the processing sequence, is the single most important step and, unfortunately, is the least well understood. Although there is an abundance of fixation mixtures recommended in the light microscopy literature, osmium tetroxide and glutaraldehyde are favored for electron microscopy. These fixatives react vigorously with proteins at the molecular level. There is clear evidence for the cross-linking of proteins both by osmium tetroxide and glutaraldehyde and cross-linking may be a necessary if not sufficient condition to define fixatives as a class.

Polishing process effect on the image of dispersed precipitates

1984 ◽  
Vol 42 ◽  
pp. 534-535
Author(s):  
C.T. Hu ◽  
C.W. Allen
Keyword(s):  
Matrix Material ◽  
Specimen Preparation ◽  
Second Phase ◽  
Precipitate Particle ◽  
Polishing Process ◽  
Second Phase Particles ◽  
The Matrix ◽  
Surface Profiles ◽  
Thinning Process

One important problem in determination of precipitate particle size is the effect of preferential thinning during TEM specimen preparation. Figure 1a schematically represents the original polydispersed Ni3Al precipitates in the Ni rich matrix. The three possible type surface profiles of TEM specimens, which result after electrolytic thinning process are illustrated in Figure 1b. c. & d. These various surface profiles could be produced by using different polishing electrolytes and conditions (i.e. temperature and electric current). The matrix-preferential-etching process causes the matrix material to be attacked much more rapidly than the second phase particles. Figure 1b indicated the result. The nonpreferential and precipitate-preferential-etching results are shown in Figures 1c and 1d respectively.

The influence of heat treatment on the fiber/matrix interface in a SiC-reinforced glassceramic

1988 ◽  
Vol 46 ◽  
pp. 742-743
Author(s):  
E. Bischoff ◽  
O. Sbaizero
Keyword(s):  
Heat Treatment ◽  
Aluminum Silicate ◽  
Lithium Aluminum ◽  
Interfacial Region ◽  
Specimen Preparation ◽  
Crack Wake ◽  
Pull Out ◽  
Annealed Material ◽  
Ultrasonic Drilling ◽  
Fiber Matrix Interface

Fiber or whisker reinforced ceramics show improved toughness and strength. Bridging by intact fibers in the crack wake and fiber pull-out after failure contribute to the additional toughness. These processes are strongly influenced by the sliding and debonding resistance of the interfacial region. The present study examines the interface in a laminated 0/90 composite consisting of SiC (Nicalon) fibers in a lithium-aluminum-silicate (LAS) glass-ceramic matrix. The material shows systematic changes in sliding resistance upon heat treatment.As-processed samples were annealed in air at 800 °C for 2, 4, 8, 16 and 100 h, and for comparison, in helium at 800 °C for 4 h. TEM specimen preparation of as processed and annealed material was performed with special care by cutting along directions having the fibers normal and parallel to the section plane, ultrasonic drilling, dimpling to 100 pm and final ionthinning. The specimen were lightly coated with Carbon and examined in an analytical TEM operated at 200 kV.

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