Laparoscopic Segmentectomy IV Using Hepatic Round Ligament Approach Combined with Fluorescent Negative Staining Method

Utilization of electron microscopy possessing a high resolution enriched the picture of macromolecular organization of chloroplasts, their individual structural elements and molecular complexes. Great possibilities have been opened in this direction with the development of negative staining method and other most recent ones allowing to study macromolecular topography of photosynthetic membranes and producing out of them grana thylakoids and stroma thylakoids. we succeeded in isolation of stroma thylakoids by means of developing the conditions of fragmentation of isolated chloroplasts. They differed radically from grana thylakoids in form and size. The analysis of submicroscopic and macromolecular organization of thylakoids was carried out by the method of negative staining (2% PTA) and ultrathin sections. Grana thylakoids (Fig. 1) isolated from chloroplasts with the aid of high pressure (150 atm) were observed in the form of rounded membranes with the diameter about 0. 5 Mm and had the particles of coupling factor on their surface (about 100 Å in diameter).

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Negative staining versus rotational evaporation

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100076597 ◽

1983 ◽

Vol 41 ◽

pp. 598-599

Author(s):

H. Schmiady ◽

C. Kreuzfeldt ◽

E. Reuber ◽

B. Tesche

Keyword(s):

Test Specimen ◽

Staining Method ◽

Three Dimensional ◽

Ribosomal Subunit ◽

Negative Staining ◽

Positive Staining ◽

Negative Stain ◽

40S Ribosomal Subunit ◽

Dimensional Reconstruction ◽

Tilt Series

This comparative study demonstrates the salient differences between the two most common methods of contrasting subcellular particles, using the 40S ribosomal subunit from saccharomyces cerevisiae as test specimen because its socalled beak-like L (left) and R (right) particle projections can be unequivocally distinguished.The negative staining method produces images which are not artifact-free because it is a combination of fixation and chemical staining and, therefore, not neutral towards the object. For three-dimensional reconstruction it is unsatisfactory because:(1) The affinity of the support film to the negative stain solution differs so greatly that sometimes undesired positive staining effects occur.(2) The rendition of the structure depends on the embedding--that is, on the level of the negative stain; fluctuations in the level lead to a loss and/or change of the structure, and thus to difficulties in interpreting the stain distribution, especially when reconstruction of the object by means of tilt series is planned.

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Comparative study of ultrastructure of HIV and SIV

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100159254 ◽

1990 ◽

Vol 48 (3) ◽

pp. 342-343

Author(s):

Takashi Nakano ◽

Shunro Imura ◽

Masuyo Nakai

Keyword(s):

Osmium Tetroxide ◽

Staining Method ◽

Culture Fluid ◽

Ultrastructural Feature ◽

Uranyl Acetate ◽

Negative Staining ◽

Thin Sections ◽

Viral Particles ◽

Infected Cells ◽

Hiv 1

The ultrastructural feature of AIDS viruses (HIV and SIV) was observed by ultrathin-sectioning and negative staining method, and the surface structures of these viruses were compared.HIV-1 group including, HTLV-III and LAV-1 were used. And in HIV-2 group, LAV-2, which was isolated from a Senegalese AIDS patient, and GH-1, which was isolated from a Ghanan AIDS patient, were used. SIV group was used AGM-1, which was isolated from an African green monkey. The virions were used which was produced from Molt-4 cells after the infection of each viruses of these strains. The infected cells and viral particles, which were collected by centrifugation were fixed with 2% glutaraldehyde (GA), washed with PBS, and postfixed with 1% osmium tetroxide. The specimens were dehydrated in graded ethanol, and embedded in Epon 812. The thin sections were stained with uranyl acetate and lead citrate. On the other hand, for negative staining, 25% GA was added to the supernatant culture fluid and finally concentrated on 2% GA. Then virions were collected by ultracentrifugation.

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Ultrastructure of Glomerular Basement Membrane in Active Heymann Nephritis Rats Revealed by Tissue-Negative Staining Method

American Journal of Nephrology ◽

10.1159/000046257 ◽

2001 ◽

Vol 21 (3) ◽

pp. 249-255 ◽

Cited By ~ 1

Author(s):

Yoshiko Hayashi ◽

Kazue Hironaka ◽

Kenichi Shikata ◽

Saeko Ogawa ◽

Kosuke Ota ◽

...

Keyword(s):

Basement Membrane ◽

Glomerular Basement Membrane ◽

Staining Method ◽

Negative Staining ◽

Heymann Nephritis

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A Rapid Method for Viral Particle Detection in Viral-Induced Gastroenteritis: A TEM Study

Microscopy and Microanalysis ◽

10.1017/s143192769511185x ◽

1995 ◽

Vol 1 (5) ◽

pp. 185-189

Author(s):

M. John Hicks ◽

James P. Barrish ◽

Elizabeth S. Hayes ◽

Laurie C. Leer ◽

Mary K. Estes ◽

...

Keyword(s):

Staining Method ◽

Pediatric Population ◽

Staining Technique ◽

Negative Staining ◽

Viral Gastroenteritis ◽

Particle Detection ◽

Fecal Samples ◽

Infectious Gastroenteritis ◽

Viral Particles ◽

Tem Method

Infectious gastroenteritis is a common cause of hospitalization in the pediatric population. The most frequent cause of gastroenteritis is viral in origin. The purpose of this study was to compare a rapid modified negative-staining TEM method with the conventional pseudoreplica technique in detection of viral particles in fecal samples from children with viral gastroenteritis. The modified negative-staining method resulted in a significantly higher (2.5 ± 0.5, p = 0.02) viral rating score than that for the conventional pseudoreplica technique (1.7 ± 0.4). In addition, the preparation time for the negative-staining method was approximately one fifth that for the conventional pseudoreplica technique. Rapid diagnosis of viral gastroenteritis may be made by ultrastructural detection of viral particles in fecal samples using the negative staining technique.

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