Electron Microscopic Demonstration of Adenovirus in Appendix Vermiformis in a Case of Ileocecal Intussusception

PEDIATRICS ◽  
1973 ◽  
Vol 51 (3) ◽  
pp. 566-570
Author(s):  
Eduardo J. Yunis ◽  
Yoshie Hashida
Keyword(s):  
Electron Microscopy ◽  
Electron Microscopic ◽  
Simple Method ◽  
Appendix Vermiformis ◽  
Viral Particles ◽  
History Of ◽  
Ileocecal Intussusception ◽  
Electron Microscopic Demonstration ◽  
Hematoxylin Eosin ◽  
Selection Of

Viral particles identical to those of adenovirus were demonstrated in the intranuclear inclusion bodies of the epithelial cells of an appendix vermiformis removed from a boy with a recent history of intussusception. Viral particles were demonstrated by electron microscopy following reprocessing of a formaldehyde-fixed, paraffin embedded and hematoxylin-eosin stained section. This simple method permits precise selection of the tissue for electron microscopy and in instances such as this gives sufficient tissue preservation to demonstrate viral particles. Our findings support the idea that some cases of intussusception may be the result of the adenovirus infection.

scholarly journals Identification of Tomato Infecting Viruses That Co-Isolate with Nanovesicles Using a Combined Proteomics and Electron-Microscopic Approach

Nanomaterials ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1922
Author(s):  
Ramila Mammadova ◽  
Immacolata Fiume ◽  
Ramesh Bokka ◽  
Veronika Kralj-Iglič ◽  
Darja Božič ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Mosaic Virus ◽  
Protein Identification ◽  
Plant Viruses ◽  
Size Exclusion ◽  
Tomato Mosaic Virus ◽  
High Yield ◽  
Plant Resources ◽  
Electron Microscopic ◽  
Viral Particles

Plant-derived nanovesicles (NVs) have attracted interest due to their anti-inflammatory, anticancer and antioxidative properties and their efficient uptake by human intestinal epithelial cells. Previously we showed that tomato (Solanum lycopersicum L.) fruit is one of the interesting plant resources from which NVs can be obtained at a high yield. In the course of the isolation of NVs from different batches of tomatoes, using the established differential ultracentrifugation or size-exclusion chromatography methods, we occasionally observed the co-isolation of viral particles. Density gradient ultracentrifugation (gUC), using sucrose or iodixanol gradient materials, turned out to be efficient in the separation of NVs from the viral particles. We applied cryogenic transmission electron microscopy (cryo-TEM), scanning electron microscopy (SEM) for the morphological assessment and LC–MS/MS-based proteomics for the protein identification of the gradient fractions. Cryo-TEM showed that a low-density gUC fraction was enriched in membrane-enclosed NVs, while the high-density fractions were rich in rod-shaped objects. Mass spectrometry–based proteomic analysis identified capsid proteins of tomato brown rugose fruit virus, tomato mosaic virus and tomato mottle mosaic virus. In another batch of tomatoes, we isolated tomato spotted wilt virus, potato virus Y and southern tomato virus in the vesicle sample. Our results show the frequent co-isolation of plant viruses with NVs and the utility of the combination of cryo-TEM, SEM and proteomics in the detection of possible viral contamination.

scholarly journals Formaldehyde as a Fixative for Light and Electron Microscopy

2000 ◽  
Vol 8 (5) ◽  
pp. 30-31
Author(s):  
Freida L. Carson
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Light And Electron Microscopy ◽  
Parallel Study ◽  
Electron Microscopic ◽  
Dual Purpose ◽  
Ultrastructural Morphology ◽  
The World ◽  
Selection Of

Since Blum discovered its hardening properties in 1893, formaldehyde has become the most widely used fixative in the world for specimens to be examined by light microscopy. However, since most commercial preparations of formaldehyde contain methanol, a protein precipitant, formaldehyde has been considered an unsatisfactory fixative for tissues to be examined by electron microscopy. In 1973, Carson et al., described a parallel study comparing the electron microscopic results of fixation with paraformaidehyde vs. formaldehyde. They found that there was no difference in the preservation of ultrastructural morphology provided that the buffer systems were identical. In 1976, McDowell and Trump described a fixative combining commercial formaldehyde and glutaraldehyde (4CF-1G). Both of these fixatives are dual purpose fixatives and preclude the selection of tissue for electron microscopy prior to fixation. They can both be prepared in large quantities and used for routine surgical specimens. The fixative containing formaldehyde alone does not need to be refrigerated and is stable for months; whereas, the formaldehyde-glutaraldehyde mixture should be refrigerated.

scholarly journals Fluorescence photooxidation with eosin: a method for high resolution immunolocalization and in situ hybridization detection for light and electron microscopy.

1994 ◽  
Vol 126 (4) ◽  
pp. 901-910 ◽  
Author(s):  
T J Deerinck ◽  
M E Martone ◽  
V Lev-Ram ◽  
D P Green ◽  
R Y Tsien ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
In Situ Hybridization ◽  
High Resolution ◽  
Light And Electron Microscopy ◽  
Three Dimensional ◽  
Substantial Improvement ◽  
Electron Microscopic ◽  
Simple Method ◽  
High Voltage Electron Microscopy

A simple method is described for high-resolution light and electron microscopic immunolocalization of proteins in cells and tissues by immunofluorescence and subsequent photooxidation of diaminobenzidine tetrahydrochloride into an insoluble osmiophilic polymer. By using eosin as the fluorescent marker, a substantial improvement in sensitivity is achieved in the photooxidation process over other conventional fluorescent compounds. The technique allows for precise correlative immunolocalization studies on the same sample using fluorescence, transmitted light and electron microscopy. Furthermore, because eosin is smaller in size than other conventional markers, this method results in improved penetration of labeling reagents compared to gold or enzyme based procedures. The improved penetration allows for three-dimensional immunolocalization using high voltage electron microscopy. Fluorescence photooxidation can also be used for high resolution light and electron microscopic localization of specific nucleic acid sequences by in situ hybridization utilizing biotinylated probes followed by an eosin-streptavidin conjugate.

scholarly journals APEX-Gold: A genetically-encoded particulate marker for robust 3D electron microscopy

2020 ◽  
Author(s):  
James Rae ◽  
Charles Ferguson ◽  
Nicholas Ariotti ◽  
Richard I. Webb ◽  
Han-Hao Cheng ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Electron Tomography ◽  
Ion Beam ◽  
Expression System ◽  
Internal Standard ◽  
Freeze Substitution ◽  
Ultrastructural Localization ◽  
Cytoskeletal Proteins ◽  
Electron Microscopic ◽  
Simple Method

AbstractGenetic tags allow rapid localization of tagged proteins in cells and tissues. APEX, an ascorbate peroxidase, has proven to be one of the most versatile and robust genetic tags for ultrastructural localization by electron microscopy. Here we describe a simple method, APEX-Gold, which converts the diffuse oxidized diaminobenzidine reaction product of APEX into a silver/gold particle akin to that used for immunogold labelling. The method increases the signal to noise ratio for EM detection, providing unambiguous detection of the tagged protein, and creates a readily quantifiable particulate signal. We demonstrate the wide applicability of this method for detection of membrane proteins, cytoplasmic proteins and cytoskeletal proteins. The method can be combined with different electron microscopic techniques including fast freezing and freeze substitution, focussed ion beam scanning electron microscopy, and electron tomography. The method allows detection of endogenously expressed proteins in genome-edited cells. We make use of a cell-free expression system to generate membrane particles with a defined quantum of an APEX-fusion protein. These particles can be added to cells to provide an internal standard for estimating absolute density of expressed APEX-fusion proteins.

Electron Microscopic Observations of a Human Brain Abscess

1973 ◽  
Vol 31 ◽  
pp. 392-393
Author(s):  
Sant S. Sekhon
Keyword(s):  
Osmium Tetroxide ◽  
Viral Encephalitis ◽  
Cell Injury ◽  
Brain Biopsy ◽  
Uranyl Acetate ◽  
Temporal Region ◽  
Electron Microscopic ◽  
Viral Particles ◽  
Grand Mal ◽  
History Of

A brain biopsy taken from an abscess located in the left temporal region of a 55-year-old caucasian male provided the basis for the present observations. The patient had a history of viral encephalitis, and a chronic brain syndrome secondary to this. He also had a positive history of grand mal seizuies probably from the viral encephalitis. The main objectives of this study were to demonstrate if possible the presence of viral particles in nerve cells and to observe and describe ultrastructural alterations and other morphological expressions of cell injury.Brain tissue from the abscess area was fixed in 2.5% cacodylate-buffered glutaraldehyde-formaldehyde mixture and post-fixed in 1% cacodylate-buffered osmium tetroxide, dehydrated and embedded in Araldite. The sections were stained with a combination of uranyl acetate and lead citrate and examined under RCA EMU 3E electron microscope.

Ruthenium red-osmium bridging with TCH: new technique to stain biological specimens for light and TEM and to coat them for SEM

1988 ◽  
Vol 46 ◽  
pp. 20-21
Author(s):  
B. Giammara ◽  
J. Hanker
Keyword(s):  
Electron Microscopy ◽  
Microscopic Study ◽  
Osmium Tetroxide ◽  
Light And Electron Microscopy ◽  
Ruthenium Red ◽  
Surface Coat ◽  
Repeated Application ◽  
Electron Microscopic ◽  
Sputter Coating ◽  
Selection Of

The demonstration and coating of glycomacromolecular surface coat components of biological specimens (1) with ruthenium red (RR, Fig. 1) is improved by treating with osmium tetroxide (2) probably due to its attachment to glycolipids. Since 1966 studies have shown how bridging osmium to osmium with thiocarbohydrazide (TCH, Fig. 2) can result in improvement in contrast of biological specimens (3,4) for light and electron microscopy. Since 1973 this bridging procedure has widely been applied (5,6) to obtain a conductive coating for biological specimens for SEM eliminating the need for sputter coating. Improvement in conductance of uncoated specimens for EM has also been obtained (6) by bridging osmium with p-phenylenediamine hydrochloride (PPD). The improvement in conductance of RR coated biological specimens for SEM by OsO4 treatment without TCH (2) required repeated application of the reagent solutions and did not result in sufficient staining of the glycomacromolecules and glycolipids for the light microscopic selection of areas for electron microscopic study.

scholarly journals Light and electron microscopic demonstration of sialic acid residues with the lectin from Limax flavus: a cytochemical affinity technique with the use of fetuin-gold complexes.

1984 ◽  
Vol 32 (11) ◽  
pp. 1167-1176 ◽  
Author(s):  
J Roth ◽  
J M Lucocq ◽  
P M Charest
Keyword(s):  
Electron Microscopy ◽  
Sialic Acid ◽  
Light And Electron Microscopy ◽  
Electron Microscopic ◽  
Gold Complexes ◽  
Thin Sections ◽  
Tissue Sections ◽  
Lowicryl K4m ◽  
Hydrolysis Of ◽  
Electron Microscopic Demonstration

The development of a cytochemical affinity technique for the demonstration of sialic acid residues by light and electron microscopy is reported. The lectin from the slug Limax flavus, with its narrow specificity for N-acetyl- and N-glycolylneuraminic acid, was applied to tissue sections. Subsequently fetuin-gold complexes were used to visualize the tissue-bound lectin. Different cytochemical controls, including sugar inhibition tests, neuraminidase digestion, the use of fetuin-gold complexes alone, or acid hydrolysis of sections, proved the specificity of the technique. Postembedding staining was performed on frozen, paraffin, or semithin resin sections for light microscopy and on thin sections from low temperature Lowicryl K4M-embedded material for electron microscopy. The distribution of sialic acid residues in rat pancreas, liver, and colonic mucosa was investigated.

scholarly journals A SEMIAUTOMATIC INSTRUMENT FOR THE RADIOAUTOGRAPHIC COATING TECHNIQUE

1966 ◽  
Vol 14 (12) ◽  
pp. 923-928 ◽  
Author(s):  
BEATRIX MARKUS KOPRIWA
Keyword(s):  
Silver Bromide ◽  
Electron Microscopic ◽  
Simple Method ◽  
Nuclear Track Emulsion ◽  
Withdrawal Speed ◽  
Emulsion Layer ◽  
Semiautomatic Instrument ◽  
Mechanical Coating ◽  
Electron Microscopic Radioautography ◽  
Selection Of

By means of a mechanical coating instrument a fast, simple method to coat specimens with liquid nuclear track emulsion has been devised for quantitative light and electron microscopic radioautography. In both cases, the section is mounted on a glass slide. After the vertically held slide has been immersed in the melted emulsion, the instrument withdraws it at a slow, constant speed. As a result, the specimen is coated with a thin, uniform emulsion layer composed of homogeneously distributed silver bromide crystals. The thickness of the emulsion coat may be standardized by selection of an optimal combination of emulsion dilution, temperature and withdrawal speed.

scholarly journals Electron microscopic demonstration of neural connections using horseradish peroxidase: a comparison of the tetramethylbenzidine procedure with seven other histochemical methods.

1982 ◽  
Vol 30 (5) ◽  
pp. 425-435 ◽  
Author(s):  
K A Carson ◽  
M M Mesulam
Keyword(s):  
Electron Microscopy ◽  
Spinal Cord ◽  
Motor Neurons ◽  
Lumbar Spinal Cord ◽  
Reaction Products ◽  
Electron Microscopic ◽  
Anterograde Transport ◽  
Microscopic Demonstration ◽  
Lumbar Spinal ◽  
Electron Microscopic Demonstration

Eight methods for the electron microscopic demonstration of horseradish peroxidase (HRP) labeling have been compared in adjacent series of vibratome sections of mouse lumbar spinal cord. The tracer, a HRP-wheat germ agglutinin (WGA) conjugate, was injected into the gastrocnemius muscle complex. Following retrograde axonal transport to the lumbar motor neurons and transganglionic anterograde transport of the tracer to the dorsal horn, the HRP activity was demonstrated in eight series of adjacent sections of lumbar spinal cord using eight methods. These included procedures using tetramethylbenzidine (TMB), benzidine dihydrochloride (BDHC), o-tolidine, paraphenylenediamine-pyrocatechol (PPD-PC), and 4 methods using 3,3'-diaminobenzidine (DAB). All eight methods were able to demonstrate both retrograde labeling of motor neurons and transganglionic anterograde transport into the dorsal horn. However, there were differences in the appearance of the various reaction products under the electron microscope. In addition, differences in the distribution of the reaction products were observed by both light and electron microscopy. The largest distribution of reaction product was observed with TMB. BDHC and o-tolidine were next, followed by the DAB procedures and PPD-PC. The TMB, BDHC, and o-tolidine reaction products were all found to be suitable for electron microscopy. The TMB reaction product was electron dense and had a very distinctive crystalloid appearance that made identification of HRP-labeled neuronal profiles easy and unequivocal.

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