Digestion techniques for exposure of smooth muscle cells in cerebral arterioles

1989 ◽  
Vol 47 ◽  
pp. 942-943
Author(s):  
S. Ghoneim ◽  
G. Baumbach
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Smooth Muscle ◽  
Basement Membrane ◽  
Three Dimensional ◽  
Type Ii ◽  
Sprague Dawley ◽  
Packed Red Blood Cells ◽  
Cerebral Arterioles ◽  
Scanning Electron

Scanning electron microscopy has proven to be a useful tool for the quantitation of three- dimensional characteristics of vascular smooth muscle. A major impediment to obtaining useful preparations of cerebral blood vessels for scanning electron microscopy is that smooth muscle in the tunica media is covered by a thin layer of basement membrane and by arachnoid tissue which contains collagen. The goal of this study was to develop a reliable, reproducible method for removing basement membrane and arachnoid tissue from cerebral arterioles, while at the same time leaving the smooth muscle intact.We compared several methods of tissue digestion in pial arterioles in adult Sprague-Dawley rats. The cerebral circulation was perfused with glutaraldehyde (2.5%) in cacodylate buffer (0.1 M) via the ascending aorta and then reinfused with packed red blood cells. In one group of rats, whole brains were immersed in osmium tetroxide (2%) for 2 hours (1 hr at 5°C and 1 hr at 40°C), followed by immersion in HC1 (8 N at 60°C) for 20-25 minutes. In another group, whole brains were dipped briefly in HC1 (8 N at 60°C) and then immersed in collagenase type II (2 mg/ml at 37° C; Sigma) for 12 hours. In a third group of rats, individual arterioles were dissected from the brain with a microsurgical knife, dipped briefly in HC1 (5-10 seconds) or KOH (5M at 60°C for 2-3minutes), and immersed in collagenase type II alone (12 hours), or in combination with collagenase type IV (2 mg/ml; 6-12 hours) and/or pepsin (2 mg/ml; 6-12 hours).

Scanning Electron Microscopy of the Polycystic Ovary in Constant Light Rats

1981 ◽  
Vol 39 ◽  
pp. 538-539
Author(s):  
K. B. Singh ◽  
F. K. Khosho
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Polycystic Ovary ◽  
Three Dimensional ◽  
Continuous Light ◽  
Female Rats ◽  
Constant Light ◽  
Sprague Dawley ◽  
Polycystic Ovaries ◽  
Scanning Electron

Exposure of female rats to continuous light eventually induces persistent estrus (PE), a syndrome which in many respects resembles the human polycystic ovary syndrome. Light microscopic studies on the polycystic ovaries of constant light PE rats have been previously reported. In the present study, we have utilized scanning electron microscopy (SEM) to provide information regarding three-dimensional changes occurring on the surface of the polycystic ovary.Following exposure to continuous light for 50 days, PE was induced in a group of Sprague-Dawley female rats according to the procedure described elsewhere (2). Polycystic ovaries from constant light PE rats and ovaries from controls were fixed in 2.5% glutaraldehyde in 0.1M Na cacodylate buffer, pH 7.3 at room temperature for 24 hours. They were then cut into two equal halves along their long axis and fixed for an additional 3-4 hours. The specimens were processed for SEM studies by a modified OTOTO technique, dehydrated and critical point dried with CO2.

scholarly journals Microscopic study of edema in hydatidiform mole

2014 ◽  
Vol 14 (3) ◽  
pp. 261-268
Author(s):  
Olivar C. Castejón ◽  
Aury Caraballo ◽  
Oliver Castejón ◽  
Elizabeth Cedeño
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Basement Membrane ◽  
Light Microscopy ◽  
Hydatidiform Mole ◽  
Three Dimensional ◽  
Wide Range ◽  
Observation Protocol ◽  
Microscopy Techniques ◽  
Scanning Electron

Objectives: the purpose of this study is to use light microscopy and scanning electron microscopy to determine the effect of edema on the structure of the molar vesicle. Methods: samples were taken from the complete hydatidiform mole and processed using conventional light and scanning electron microscopy techniques and an observation protocol that identified four variables: factors underlying the development of edema; the condition of the trophoblast basement membrane, development of the villi, accumulation and degeneration of sulphated mucosubstances at stromal level. Results: light microscopy showed a permeable trophoblastic basement membrane, a swollen syncytium, edematous regions disorganizating the stromal region and causing ischemic necrosis of cells. Using scanning electron microscopy, the basement membrane was found to be distended and thickened, with large irregular holes for the entry and movement of liquid, leaving a wide range of fluids during the influx process and depriving stromal cells of nutrition. Conclusions: a new three-dimensional view of the changes brought about by the entry of fluids into the stroma of molar hydropic vesicles was provided by scanning electron microscopy and confirmed by light microscopy, thereby explaining the changes occurring at the level of the stroma as an effect of the edema.

Scanning and Transmission Microscopy of Nematocyst Batteries in Epitheliomuscular Cells of Hydra

1971 ◽  
Vol 29 ◽  
pp. 410-411 ◽  
Author(s):  
Jane A. Westfall ◽  
S. Yamataka ◽  
Paul D. Enos
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Osmium Tetroxide ◽  
External Surface ◽  
Three Dimensional ◽  
Surface Structures ◽  
Transmission Microscopy ◽  
Transmission Electron ◽  
Freeze Dried ◽  
Scanning Electron

Scanning electron microscopy (SEM) provides three dimensional details of external surface structures and supplements ultrastructural information provided by transmission electron microscopy (TEM). Animals composed of watery jellylike tissues such as hydras and other coelenterates have not been considered suitable for SEM studies because of the difficulty in preserving such organisms in a normal state. This study demonstrates 1) the successful use of SEM on such tissue, and 2) the unique arrangement of batteries of nematocysts within large epitheliomuscular cells on tentacles of Hydra littoralis.Whole specimens of Hydra were prepared for SEM (Figs. 1 and 2) by the fix, freeze-dry, coat technique of Small and Màrszalek. The specimens were fixed in osmium tetroxide and mercuric chloride, freeze-dried in vacuo on a prechilled 1 Kg brass block, and coated with gold-palladium. Tissues for TEM (Figs. 3 and 4) were fixed in glutaraldehyde followed by osmium tetroxide. Scanning micrographs were taken on a Cambridge Stereoscan Mark II A microscope at 10 KV and transmission micrographs were taken on an RCA EMU 3G microscope (Fig. 3) or on a Hitachi HU 11B microscope (Fig. 4).

Scanning Electron Microscopy of Human Jejunum in Whipple's Disease

1977 ◽  
Vol 35 ◽  
pp. 354-355
Author(s):  
John H. L. Watson ◽  
C. N. Sun
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Basement Membrane ◽  
Lamina Propria ◽  
Brush Border ◽  
Whipple’S Disease ◽  
Whipple's Disease ◽  
Biopsy Specimens ◽  
Brush Borders ◽  
Scanning Electron

That the etiology of Whipple's disease could be bacterial was first suggested from electron micrographs in 1960. Evidence for binary fission of the bacteria, their phagocytosis by histiocytes in the lamina propria, their occurrence between and within the cells of the epithelium and on the brush border of the lumen were reported later. Scanning electron microscopy has been applied by us in an attempt to confirm the earlier observations by the new technique and to describe the bacterium further. Both transmission and scanning electron microscopy have been used concurrently to study the same biopsy specimens, and transmission observations have been used to confirm those made by scanning.The locations of the brush borders, the columnar epithelial cells, the basement membrane and the lamina propria beneath it were each easily identified by scanning electron microscopy. The lamina propria was completely filled with the wiener-shaped bacteria, Fig. 1.

Scanning Electron Microscopy and Electron Microprobe Analysis of Malignant Cell Cultures

1968 ◽  
Vol 26 ◽  
pp. 164-165
Author(s):  
R. I. Johnsson-Hegyeli ◽  
A. F. Hegyeli ◽  
D. K. Landstrom ◽  
W. C. Lane
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Cell Cultures ◽  
Electron Microprobe ◽  
Microprobe Analysis ◽  
Electron Microprobe Analysis ◽  
Three Dimensional ◽  
Malignant Cell ◽  
Interference Microscopy ◽  
Scanning Electron

Last year we reported on the use of reflected light interference microscopy (RLIM) for the direct color photography of the surfaces of living normal and malignant cell cultures without the use of replicas, fixatives, or stains. The surface topography of living cells was found to follow underlying cellular structures such as nuceloli, nuclear membranes, and cytoplasmic organelles, making possible the study of their three-dimensional relationships in time. The technique makes possible the direct examination of cells grown on opaque as well as transparent surfaces. The successful in situ electron microprobe analysis of the elemental composition and distribution within single tissue culture cells was also reported.This paper deals with the parallel and combined use of scanning electron microscopy (SEM) and the two previous techniques in a study of living and fixed cancer cells. All three studies can be carried out consecutively on the same experimental specimens without disturbing the cells or their structural relationships to each other and the surface on which they are grown. KB carcinoma cells were grown on glass coverslips in closed Leighto tubes as previously described. The cultures were photographed alive by means of RLIM, then fixed with a fixative modified from Sabatini, et al (1963).

scholarly journals Three-dimensional relationships between tumor cells and microcirculation with double cyanine immunolabeling, laser scanning confocal microscopy, and computer-assisted reconstruction: an alternative to cast corrosion preparations.

1994 ◽  
Vol 42 (5) ◽  
pp. 681-686 ◽  
Author(s):  
V Rummelt ◽  
L M Gardner ◽  
R Folberg ◽  
S Beck ◽  
B Knosp ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Blood Vessels ◽  
Tumor Cells ◽  
Laser Scanning ◽  
Three Dimensional ◽  
Melanoma Cells ◽  
Tumor Blood Vessels ◽  
The Relationship ◽  
Scanning Electron

The morphology of the microcirculation of uveal melanomas is a reliable market of tumor progression. Scanning electron microscopy of cast corrosion preparations can generate three-dimensional views of these vascular patterns, but this technique sacrifices the tumor parenchyma. Formalin-fixed wet tissue sections 100-150 microns thick from uveal melanomas were stained with the lectin Ulex europaeus agglutinin I (UEAI) and proliferating cell nuclear antigen (PCNA) to demonstrate simultaneously the tumor blood vessels and proliferating tumor cells. Indocarbocyanine (Cy3) was used as a fluorophore for UEAI and indodicarbocyanine (Cy5) was used for PCNA. Double labeled sections were examined with a laser scanning confocal microscope. Images of both stains were digitized at the same 5-microns intervals and each of the two images per interval was combined digitally to form one image. These combined images were visualized through voxel processing to study the relationship between melanoma cells expressing PCNA and various microcirculatory patterns. This technique produces images comparable to scanning electron microscopy of cast corrosion preparations while permitting simultaneous localization of melanoma cells expressing PCNA. The microcirculatory tree can be viewed from any perspective and the relationship between tumor cells and the tumor blood vessels can be studied concurrently in three dimensions. This technique is an alternative to cast corrosion preparations.

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