Methods for making stereoslides and -prints, with freeze-etched olfactory epithelium as example

Author(s):  
Bert Ph. M. Menco ◽  
Ido F. Menco ◽  
Frans L.T. Verdonk

Previously we presented an extensive study of the distributions of intramembranous particles of structures in apical surfaces of nasal olfactory and respiratory epithelia of the Sprague-Dawley rat. For the same structures these distributions were compared in samples which were i) chemically fixed and cryo-protected with glycerol before cryo-fixation, after excision, and ii)ultra-rapidly frozen by means of the slam-freezing method. Since a three-dimensional presentation markedly improves visualization of structural features micrographs were presented as stereopairs. Two exposures were made by tiling the sample stage of the electron microscope 6° in either direction with an eucentric goniometer. The negatives (Agfa Pan 25 Professional) were reversed with Kodak Technical Pan Film 2415 developed in D76 1:1. The prints were made from these reversed negatives. As an example tight-junctional features of an olfactory supporting cell in a region where this cell conjoined with two other cells are presented (Fig. 1).

Author(s):  
Takashi Ichijo ◽  
Yasuo Yamashita ◽  
tatsuo Terashima ◽  
Humio Nagata ◽  
Sadahiko Okamura ◽  
...  

Recently it has become generally agreed that tooth and bone crystals consist principally of hydroxyapatite, Ca10(PO4)6(OH)2. Over several years, we have been pursuing observations on apatite crystals composing the teeth and bones at the level of their atomic configuration using transmission electron microscope for the purpose of examining their structural features of biological crystals. In this study, observations using the same approach were conducted a higher resolution than before. In this report, because of the highest resolution ever obtained that, crystals could be observed at a level of ⅙ lattice (0.136 nm intervals). Using this approach, subsequentry, the possibility of the three-dimensional observation on the configration composing hydroxyapatite structure in the unit cells was pursued. The crystals composing teeth and bone known to have slight curves and twists. The authors paid attention to this fact, and attempted to three-dimensionally observe the portion where the c axis of the crystal unit cells tilt slightly due to some proper curving of the crystals.


Author(s):  
O.C. de Hodgins ◽  
K. R. Lawless ◽  
R. Anderson

Commercial polyimide films have shown to be homogeneous on a scale of 5 to 200 nm. The observation of Skybond (SKB) 705 and PI5878 was carried out by using a Philips 400, 120 KeV STEM. The objective was to elucidate the structural features of the polymeric samples. The specimens were spun and cured at stepped temperatures in an inert atmosphere and cooled slowly for eight hours. TEM micrographs showed heterogeneities (or nodular structures) generally on a scale of 100 nm for PI5878 and approximately 40 nm for SKB 705, present in large volume fractions of both specimens. See Figures 1 and 2. It is possible that the nodulus observed may be associated with surface effects and the structure of the polymers be regarded as random amorphous arrays. Diffraction patterns of the matrix and the nodular areas showed different amorphous ring patterns in both materials. The specimens were viewed in both bright and dark fields using a high resolution electron microscope which provided magnifications of 100,000X or more on the photographic plates if desired.


Author(s):  
L. V. Leak

Electron microscopic observations of freeze-fracture replicas of Anabaena cells obtained by the procedures described by Bullivant and Ames (J. Cell Biol., 1966) indicate that the frozen cells are fractured in many different planes. This fracturing or cleaving along various planes allows one to gain a three dimensional relation of the cellular components as a result of such a manipulation. When replicas that are obtained by the freeze-fracture method are observed in the electron microscope, cross fractures of the cell wall and membranes that comprise the photosynthetic lamellae are apparent as demonstrated in Figures 1 & 2.A large portion of the Anabaena cell is composed of undulating layers of cytoplasm that are bounded by unit membranes that comprise the photosynthetic membranes. The adjoining layers of cytoplasm are closely apposed to each other to form the photosynthetic lamellae. Occassionally the adjacent layers of cytoplasm are separated by an interspace that may vary in widths of up to several 100 mu to form intralamellar vesicles.


Author(s):  
P. Dayanandan ◽  
P. B. Kaufman

A three dimensional appreciation of the guard cell morphology coupled with ultrastjuctural studies should lead to a better understanding of their still obscure dynamics of movement. We have found the SEM of great value not only in studies of the surface details of stomata but also in resolving the structures and relationships that exist between the guard and subsidiary cells. We now report the isolation and SEM studies of guard cells from nine genera of plants.Guard cells were isolated from the following plants: Psilotum nudum, four species of Equisetum, Cycas revoluta, Ceratozamia sp., Pinus sylvestris, Ephedra cochuma, Welwitschia mirabilis, Euphorbia tirucalli and Allium cepa.


Author(s):  
J.L. Williams ◽  
K. Heathcote ◽  
E.J. Greer

High Voltage Electron Microscope already offers exciting experimental possibilities to Biologists and Materials Scientists because the increased specimen thickness allows direct observation of three dimensional structure and dynamic experiments on effectively bulk specimens. This microscope is designed to give maximum accessibility and space in the specimen region for the special stages which are required. At the same time it provides an ease of operation similar to a conventional instrument.


Author(s):  
S. Shirahama ◽  
G. C. Engle ◽  
R. M. Dutcher

A transplantable carcinoma was established in North West Sprague Dawley (NWSD) rats by use of X-irradiation by Engle and Spencer. The tumor was passaged through 63 generations over a period of 32 months. The original tumor, an adenocarcinoma, changed into an undifferentiated carcinoma following the 19th transplant. The tumor grew well in NWSD rats of either sex at various ages. It was invariably fatal, causing death of the host within 15 to 35 days following transplantation.Tumor, thymus, spleen, and plasma from 7 rats receiving transplants of tumor at 3 to 9 weeks of age were examined with an electron microscope at intervals of 8, 15, 22 and 30 days after transplantation. Four normal control rats of the same age were also examined. The tissues were fixed in glutaraldehyde, postfixed in osmium tetroxide and embedded in Epon. The plasma was separated from heparanized blood and processed as previously described for the tissue specimens. Sections were stained with uranyl acetate followed by lead citrate and examined with an RCA EMU-3G electron microscope.


Author(s):  
P. S. Kotval ◽  
C. J. Dewit

The structure of Ta2O5 has been described in the literature in several different crystallographic forms with varying unit cell lattice parameters. Earlier studies on films of Ta2O5 produced by anodization of tantalum have revealed structural features which are not consistent with the parameters of “bulk” Ta2O5 crystalsFilms of Ta2O5 were prepared by anodizing a well-polished surface of pure tantalum sheet. The anodic films were floated off in distilled water, collected on grids, dried and directly examined in the electron microscope. In all cases the films were found to exhibit diffraction patterns representative of an amorphous structure. Using beam heating in the electron microscope, recrystallization of the amorphous films can be accomplished as shown in Fig. 1. As suggested by earlier work, the recrystallized regions exhibit diffraction patterns which consist of hexagonal arrays of main spots together with subsidiary rows of super lattice spots which develop as recrystallization progresses (Figs. 2a and b).


2013 ◽  
Vol 19 (S5) ◽  
pp. 58-61 ◽  
Author(s):  
Mino Yang ◽  
Jun-Ho Lee ◽  
Hee-Goo Kim ◽  
Euna Kim ◽  
Young-Nam Kwon ◽  
...  

AbstractDistribution of wax in laser printer toner was observed using an ultra-high-voltage (UHV) and a medium-voltage transmission electron microscope (TEM). As the radius of the wax spans a hundred to greater than a thousand nanometers, its three-dimensional recognition via TEM requires large depth of focus (DOF) for a volumetric specimen. A tomogram with a series of the captured images would allow the determination of their spatial distribution. In this study, bright-field (BF) images acquired with UHV-TEM at a high tilt angle prevented the construction of the tomogram. Conversely, the Z-contrast images acquired by the medium-voltage TEM produced a successful tomogram. The spatial resolution for both is discussed, illustrating that the image degradation was primarily caused by beam divergence of the Z-contrast image and the combination of DOF and chromatic aberration of the BF image from the UHV-TEM.


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