Histological effects of vitamin A on limb regeneration in the larval axolotl, Ambystoma mexicanum

1990 ◽  
Vol 68 (1) ◽  
pp. 159-167 ◽  
Author(s):  
Steven R. Scadding

Vitamin A causes profound changes in the development of pattern during amphibian limb regeneration. Vitamin A effects include the induction of duplications of skeletal structures in the anteroposterior, proximodistal, and dorsoventral axes. The purpose of this investigation was to study the underlying histological changes that are induced in the regenerating limb of the larval axolotl, Ambystoma mexicanum, by treatment with vitamin A. Axolotl larvae (7–10 cm in length) had forelimbs amputated through the radius and ulna and were then immersed in 75 IU/mL retinol palmitate for 14 days. Limbs were removed and fixed at intervals over the period of regeneration, both during and beyond the period of retinoid treatment. They were then examined in the light microscope, scanning electron microscope, or transmission electron microscope. Compared with the controls, the retinol palmitate treated regenerating limbs exhibited the development of an eccentric epidermal cap which was always displaced towards the posterior edge of the limb. Beneath this epidermal cap, the density of the cells of the blastema was greater than that of the cells towards the anterior edge of the developing blastema where the cells were much less densely arranged. Epidermal changes induced by retinol palmitate included the development of a very uneven and creviced surface, with a great deal of variation in cell size, and the development of ciliated cells in the surface layer of the epidermis. The significance of these observations for pattern modification by vitamin A are discussed.

1974 ◽  
Vol 83 (1) ◽  
pp. 102-106 ◽  
Author(s):  
Tomoyuki Hoshino ◽  
Kenji Matsumoto

Using a scanning electron microscope, a group of ciliated cells was found in the area of the utricular macula of an adult cat. A cross section of the same specimen viewed with a transmission electron microscope revealed that the hairs observed had the characteristic inner structure of true cilia. As the same cell group was not found in either the contralateral ear of the same cat or in any of three other cats examined, the presence of these ciliated cells seems to be exceptional.


Author(s):  
J. D. Hutchison

When the transmission electron microscope was commercially introduced a few years ago, it was heralded as one of the most significant aids to medical research of the century. It continues to occupy that niche; however, the scanning electron microscope is gaining rapidly in relative importance as it fills the gap between conventional optical microscopy and transmission electron microscopy.IBM Boulder is conducting three major programs in cooperation with the Colorado School of Medicine. These are the study of the mechanism of failure of the prosthetic heart valve, the study of the ultrastructure of lung tissue, and the definition of the function of the cilia of the ventricular ependyma of the brain.


Author(s):  
K. Shibatomi ◽  
T. Yamanoto ◽  
H. Koike

In the observation of a thick specimen by means of a transmission electron microscope, the intensity of electrons passing through the objective lens aperture is greatly reduced. So that the image is almost invisible. In addition to this fact, it have been reported that a chromatic aberration causes the deterioration of the image contrast rather than that of the resolution. The scanning electron microscope is, however, capable of electrically amplifying the signal of the decreasing intensity, and also free from a chromatic aberration so that the deterioration of the image contrast due to the aberration can be prevented. The electrical improvement of the image quality can be carried out by using the fascionating features of the SEM, that is, the amplification of a weak in-put signal forming the image and the descriminating action of the heigh level signal of the background. This paper reports some of the experimental results about the thickness dependence of the observability and quality of the image in the case of the transmission SEM.


Author(s):  
S. Takashima ◽  
H. Hashimoto ◽  
S. Kimoto

The resolution of a conventional transmission electron microscope (TEM) deteriorates as the specimen thickness increases, because chromatic aberration of the objective lens is caused by the energy loss of electrons). In the case of a scanning electron microscope (SEM), chromatic aberration does not exist as the restrictive factor for the resolution of the transmitted electron image, for the SEM has no imageforming lens. It is not sure, however, that the equal resolution to the probe diameter can be obtained in the case of a thick specimen. To study the relation between the specimen thickness and the resolution of the trans-mitted electron image obtained by the SEM, the following experiment was carried out.


Author(s):  
Masayuki Miyoshi

In spite of various attempts, conclusive evidence to explain blood passage in the splenic red pulp does not seem to have been presented. Scanning electron microscope (SEM) observations on the rabbit spleen, originally performed by us, revealed that the sinus was lined by a perforated lattice composed of longitudinally extended rod cells and transverse cytoplasmic processes, and that perforations in the lattice were continuous to the spaces among the stellate reticulum cells of the cord. In the present study the observation was extended to the dog and rat spleens, in which the cord is more developed than in the rabbit in order to clarify the possible differences in the fine structure of the sinus wall. An attempt was also made to examine the development and distribution of macrophage in the blood passage of the red pulp.Spleens were washed and fixed by perfusion with Ringer solution and then with buffered glutaraldehyde. Small tissue cubes were dehydrated with acetone, dried in air and heated with gold. Observations were made by a JEOL SEM Type-3. One air dried tissue cube was cut into small pieces and post fixed with buffered OsO4 for examination under the transmission electron microscope (TEM).


Author(s):  
J. Temple Black

Since its introduction by Fernandez-Moran, the diamond knife has gained wide spread usage as a common material for cutting of thin sections of biological and metallic materials into thin films for examination in the transmission electron microscope. With the development of high voltage E.M. and scanning transmission E.M., microtomy applications will become increasingly important in the preparation of specimens. For those who can afford it, the diamond knife will thus continue to be an important tool to accomplish this effort until a cheaper but equally strong and sharp tool is found to replace the diamond, glass not withstanding.In Figs. 1 thru 3, a first attempt was made to examine the edge of a used (β=45°) diamond knife by means of the scanning electron microscope. Because diamond is conductive, first examination was tried without any coating of the diamond. However, the contamination at the edge caused severe charging during imaging. Next, a thin layer of carbon was deposited but charging was still extensive at high magnification - high voltage settings. Finally, the knife was given a light coating of gold-palladium which eliminated the charging and allowed high magnification micrographs to be made with reasonable resolution.


Author(s):  
J. C. Russ ◽  
E. McNatt

In order to study the retention of copper in cirrhotic liver, rats were made cirrhotic by carbon tetrachloride inhalation twice weekly for three months and fed 0.2% copper acetate ad libidum in drinking water for one month. The liver tissue was fixed in osmium, sectioned approximately 2000 Å thick, and stained with lead citrate. The section was examined in a scanning electron microscope (JEOLCO JSM-2) in the transmission electron mode.Figure 1 shows a typical area that includes a red blood cell in a sinusoid, a disse, and a portion of the cytoplasm of a hepatocyte which contains several mitochondria, peribiliary dense bodies, glycogen granules, and endoplasmic reticulum.


Author(s):  
M. K. Lamvik

When observing small objects such as cellular organelles by scanning electron microscopy, it is often valuable to use the techniques of transmission electron microscopy. The common practice of mounting and coating for SEM may not always be necessary. These possibilities are illustrated using vertebrate skeletal muscle myofibrils.Micrographs for this study were made using a Hitachi HFS-2 scanning electron microscope, with photographic recording usually done at 60 seconds per frame. The instrument was operated at 25 kV, with a specimen chamber vacuum usually better than 10-7 torr. Myofibrils were obtained from rabbit back muscle using the method of Zak et al. To show the component filaments of this contractile organelle, the myofibrils were partially disrupted by agitation in a relaxing medium. A brief centrifugation was done to clear the solution of most of the undisrupted myofibrils before a drop was placed on the grid. Standard 3 mm transmission electron microscope grids covered with thin carbon films were used in this study.


Author(s):  
Shaopeng Hu ◽  
Jianhua Wang ◽  
Zhen Li ◽  
Huei Chen ◽  
Fei Cu ◽  
...  

Gastritis from returning bile is a common disease, but the reason for the disease is not clear. As the pathologic ultrastructure research progresses, it has drawn attention to the ultrastructural change of cells in gastric mucosa by clinical workers. We observed gastric mucosa tissues of 15 patients suffering from gastritis with a transmission electron microscope (TEM) and a scanning electron microscope (SEM). It is the first report in China that fungus exists in the lamina propria of gastric mucosa tissue. The result is as follows.The gastric mucosa tissues of 15 patients suffering from gastritis were acquired by stomachoscopy. Both TEM and SEM specimens were prepared by the usual methods. Under the TEM, the epithelial surface became higher and larger. Mitochondria of the cells were swollen and cristae were disrupted. There were vacuoles in the cells. The nucleus showed disorder, heterochromatin became darker, and nucleolae could be observed.


Author(s):  
W. P. Wergin ◽  
S. Roy ◽  
E. F. Erbe ◽  
C. A. Murphy ◽  
C. D. Pooley

Larvae of the nematode, Steinernema carpocapsae Weiser strain All, were cryofixed and freezesubstituted for 3 days in acetone containing 2% osmium tetroxide according to established procedures. Following chemical fixation, the nematodes were brought to room temperature, embedded in Spurr's medium and sectioned for observation with a Hitachi S-4100 field emission scanning electron microscope that was equipped with an Oxford CT 1500 Cryotrans System. Thin sections, about 80 nm thick, similar to those generally used in conventional transmission electron microscope (TEM) studies were mounted on copper grids and stained with uranyl acetate for 30 min and lead citrate for 5 min. Sections about 2 μm thick were also mounted and stained in a similar fashion. The grids were mounted on an Oxford grid holder, inserted into the microscope and onto a cryostage that was operated at ambient temperature. Thick and thin sections of the larvae were evaluated and photographed in the SEM at different accelerating voltages. Figs. 4 and 5 have undergone contrast conversion so that the images would resemble transmitted electron micrographs obtained with a TEM.


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