Identification of the pulmonary syndrome hantavirus by direct and colloidal gold immune Electron Microscopy

1994 ◽  
Vol 52 ◽  
pp. 274-275
Author(s):  
C. D. Humphrey ◽  
C.S. Goldsmith ◽  
L. Elliott ◽  
S.R. Zaki
Keyword(s):  
Electron Microscopy ◽  
United States ◽  
Monoclonal Antibody ◽  
Colloidal Gold ◽  
Phosphotungstic Acid ◽  
Uranyl Acetate ◽  
Hantavirus Pulmonary Syndrome ◽  
Deer Mice ◽  
Immune Electron Microscopy ◽  
Negative Stain

An outbreak of unexplained acute pulmonary syndrome with high fatality was recognized in the spring of 1993 in the southwestern United States. The cause of the illness was quickly identified serologically and genetically as a hantavirus and the disease was named hantavirus pulmonary syndrome (HPS). Recently, the virus was isolated from deer mice which had been trapped near the homes of HPS patients, and cultivated in Vero E6 cells. We identified the cultivated virus by negative-stain direct and colloidal gold immune electron microscopy (EM).Virus was extracted, clarified, and concentrated from unfixed and 0.25% glutaraldehyde fixed supernatant fluids of infected Vero E6 cells by a procedure described previously. Concentrated virus suspensions tested by direct EM were applied to glow-discharge treated formvar-carbon filmed grids, blotted, and stained with 0.5% uranyl acetate (UA) or with 2% phosphotungstic acid (PTA) pH 6.5. Virus suspensions for immune colloidal gold identification were adsorbed similarly to filmed grids but incubated for 1 hr on drops of 1:50 diluted monoclonal antibody to Prospect Hill virus nucleoprotein or with 1:50 diluted sera from HPS virus infected deer mice.

scholarly journals Complete Genome Sequences of Monongahela Hantavirus from Pennsylvania, USA

2018 ◽  
Vol 7 (11) ◽  
Author(s):  
César G. Albariño ◽  
Lisa Wiggleton Guerrero ◽  
Ayan K. Chakrabarti ◽  
Pierre E. Rollin ◽  
Stuart T. Nichol
Keyword(s):  
United States ◽  
West Virginia ◽  
Complete Genome ◽  
Clinical Case ◽  
The United States ◽  
Hantavirus Pulmonary Syndrome ◽  
Deer Mice ◽  
Genome Sequences ◽  
Complete Sequences

Monongahela hantavirus was first identified in deer mice and was later found responsible for hantavirus pulmonary syndrome cases in Pennsylvania and West Virginia in the United States. Here, we report the complete sequences of Monongahela virus S, M, and L genomic segments obtained from a fatal clinical case reported in 1997.

Phosphotungstic acid staining of latexes for transmission electron microscopy

1983 ◽  
Vol 41 ◽  
pp. 30-31
Author(s):  
O. L. Shaffer ◽  
M. S. El-Aasser ◽  
J. W. Vanderhoff
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Glass Transition ◽  
Phosphotungstic Acid ◽  
Particle Morphology ◽  
Ethyl Acrylate ◽  
Uranyl Acetate ◽  
Positive Staining ◽  
Sample Treatment ◽  
Transmission Electron

Measurement of particle size and particle morphology by transmission electron microscopy (TEM) is important in the characterization of polymer latex systems. Special sample treatment methods have been developed to permit the study of latexes that present problems, such as low-glass-transition-temperature (Tg) and electron-transparent particles. Some of these methods include hardening and staining by osmium tetroxide or bromine of latexes that contain unsaturation, negative staining by uranyl acetate, and freezing of latexes with low glass-transition temperatures.We have recently found phosphotungstic acid (PTA) to be useful in both negative and positive staining of latex particles. As a negative stain, PTA can enhance the contrast between the electron-transparent particles and the dense PTA-stained background. This has been particularly useful with latexes such as poly(butyl acrylate), poly(ethyl acrylate), poly(ethylene), and other polymers of similar electron densities. As a positive stain, PTA can also react with the surface functional groups of particles, such as hydroxyl, carboxyl, and amine groups.

Electron Microscopy of Wound Healing in Patients with Hernia

1975 ◽  
Vol 33 ◽  
pp. 352-353
Author(s):  
C.N. Sun ◽  
H.J. White ◽  
R.C. Read
Keyword(s):  
Electron Microscopy ◽  
Wound Healing ◽  
Phosphate Buffer ◽  
Osmium Tetroxide ◽  
Phosphotungstic Acid ◽  
Rectus Sheath ◽  
Uranyl Acetate ◽  
Collagen Fibrils ◽  
Lead Citrate ◽  
Native Collagen

Previously we have reported the defect of collagen fibrils from herniated rectus sheath. This presentation includes additional sections from postsurgical incisions (10 days) from both control and hernia patients. Small pieces of rectus sheath were fixed in 3% glutaraldehyde in phosphate buffer (pH 7.2) and post fixed with buffered 2% osmium tetroxide. The tissues were then dehydrated in serially increasing concentrations of alcohol and embedded in Epon 812. Sections were stained with 2.5% phosphotungstic acid or uranyl acetate and lead citrate.Previously we found that collagen fibrils from "non-herniated" rectus sheath have uniform diameters and 640 Å periodicity with seven or more intraperiodic bands resembling typical native collagen fibrils, while the fibrils from fascia obtained from patients with direct herniation show considerable variation in diameter. These variations are often found in the same individual fibers with a range from 300 Å to 3000 Å.

The Influence of Ammonium Molybdate, Silicotungstic Acid and Phosphotungstic Acid on Function and Structure of Mitochondria

1970 ◽  
Vol 28 ◽  
pp. 70-71
Author(s):  
W. J. Mergner ◽  
R. Pendergrass ◽  
B. F. Trump
Keyword(s):  
Electron Microscopy ◽  
Phosphotungstic Acid ◽  
Rat Kidney ◽  
Kinetic Studies ◽  
Ammonium Molybdate ◽  
Silicotungstic Acid ◽  
Ion Permeability ◽  
Freeze Fracturing ◽  
Kidney Mitochondria ◽  
Negative Stain

A negative stain should produce no structural change in the specimen. Our results indicate that this is not the case with at least two negative stains. There is a spectrum of changes which depends on the stain used. The negative stain also causes functional alterations which involve three basic functional characteristics of the mitochondria: 1) oxidative phosphorylation, 2) electron transport and 3) ion permeability. Rat kidney mitochondria were resuspended in the negative stain for 15 minutes (except for kinetic studies). For electron microscopy they were: 1) directly applied to grids; 2) fixed with glutaraldehyde and OSO4 and embedded in Epon; and 3) prepared by freeze fracturing. Mitochondria stained with ammonium molybdate (AM) revealed two types of profiles. They had either a dense inner compartment without penetration of negative stain displaying the commonly observed image (Fig. 3 and 4). With penetration the inner membrane was studded with spheres, but the spheres were not on discernible stalks (Fig. #5).

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