Detection of Early Cytomegalovirus Infection in Pathologial Specimens by H&E, IHC, ISH, Grocott’S and Modified Steiner Silver Stain for LM and EM: A Review

2001 ◽  
Vol 7 (S2) ◽  
pp. 178-179
Author(s):  
K. Chien ◽  
ML. Heathershaw ◽  
R.C. Heusser ◽  
H. Shiroishi ◽  
R. Gonzalez ◽  
...  
Keyword(s):  
Intranuclear Inclusions ◽  
Cmv Infection ◽  
Silver Stain ◽  
Characteristic Appearance ◽  
Tissue Sections ◽  
Viral Particles ◽  
Section Size ◽  
Clear Halo ◽  
Size Limits ◽  
Microscopic Techniques

The diagnosis of cytomegalovirus (CMV) in tissue sections is usually quite easy because of the characteristic appearance of intranuclear inclusions. At times the morphological features are equivocal, the nuclear findings may be atypical or only a single cell may be affected, thereby creating uncertainty about the diagnosis. This report reviews our experience with the various microscopic techniques that can be used to make definitive identification of CMV.EM - Visualizing the viral particles in a single or even lysed cell by routine electron microscopy can make the definite diagnosis, but section size limits the usefulness in the detection of early CMV infection (fig. 1). However, if a diagnosis can not be made with certainty by the following methods, EM can be performed on stained paraffin sections for confirmation.H&E - A Hematoxylin & Eosin stain that discloses characteristic intranuclear inclusions surrounded by a clear halo indicates CMV infection (fig.2A).

A combined pseudoreplica-immunocytochemical technique for research and diagnostic virology

1990 ◽  
Vol 48 (3) ◽  
pp. 900-901
Author(s):  
Blayne Fritz ◽  
Stanley J. Naides ◽  
Kenneth Moore
Keyword(s):  
Phosphotungstic Acid ◽  
Immunogold Labelling ◽  
Uranyl Acetate ◽  
Distilled Water ◽  
Clinical Specimens ◽  
Tissue Sections ◽  
Specimen Retrieval ◽  
Transmission Electron ◽  
Viral Particles ◽  
Diagnostic Virology

The pseudoreplica method of staining viral particles for visualization by transmission electron microscopy is a very popular technique. The ability to concentrate clinical specimens while semi-embedding viral particles makes it especially well suited for morphologic and diagnostic virology. Immunolabelling viral particles with colloidal gold is a technique frequently employed by both research and diagnostic virologists. We have characterized a procedure which provides the advantage of both by modifying and combining pseudoreplica staining and immunogold labelling.Modification of specimen retrieval and delay of staining allows us to utilize pseudoreplica processed specimens within our standard immunogold labelling protocol. In brief, we absorbed samples onto 2% agarose, added.25% Formvar and wicked dry. We then floated the Formvar-virus film onto double distilled water, added grids and retrieved with parafilm. The Formvar-virus specimens were then treated as thin tissue sections within our standard two stage immunolabelling protocol. Following completion of immunogold labelling; each grid was negatively stained with phosphotungstic acid or uranyl acetate contrast stains.

scholarly journals Deep learning-based transformation of H&E stained tissues into special stains

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Kevin de Haan ◽  
Yijie Zhang ◽  
Jonathan E. Zuckerman ◽  
Tairan Liu ◽  
Anthony E. Sisk ◽  
...  
Keyword(s):  
Visual Inspection ◽  
Kidney Diseases ◽  
Periodic Acid ◽  
Needle Core Biopsy ◽  
Silver Stain ◽  
Periodic Acid Schiff ◽  
Tissue Sections ◽  
Biopsy Tissue ◽  
Hematoxylin And Eosin

AbstractPathology is practiced by visual inspection of histochemically stained tissue slides. While the hematoxylin and eosin (H&E) stain is most commonly used, special stains can provide additional contrast to different tissue components. Here, we demonstrate the utility of supervised learning-based computational stain transformation from H&E to special stains (Masson’s Trichrome, periodic acid-Schiff and Jones silver stain) using kidney needle core biopsy tissue sections. Based on the evaluation by three renal pathologists, followed by adjudication by a fourth pathologist, we show that the generation of virtual special stains from existing H&E images improves the diagnosis of several non-neoplastic kidney diseases, sampled from 58 unique subjects (P = 0.0095). A second study found that the quality of the computationally generated special stains was statistically equivalent to those which were histochemically stained. This stain-to-stain transformation framework can improve preliminary diagnoses when additional special stains are needed, also providing significant savings in time and cost.

Salmonella enterica serovar Typhimurium bacteraemia in a puppy with canine parvoviral enteritis

2020 ◽  
Vol 8 (3) ◽  
pp. e001171
Author(s):  
Josef Hanekom ◽  
Paolo Pazzi ◽  
Yolandi Rautenbach ◽  
Alischa Henning
Keyword(s):  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Peripheral Blood Smear ◽  
Elisa Test ◽  
Canine Parvovirus ◽  
Tissue Sections ◽  
Viral Particles ◽  
Blood Smears ◽  
Serovar Typhimurium ◽  
Haemorrhagic Diarrhoea

A 12-week-old female intact, pit bull terrier cross breed puppy presented with vomiting and haemorrhagic diarrhoea. Phagocytosed bacterial rods were observed on peripheral and central blood smears. A commercially available canine parvovirus ELISA test and subsequent electron microscopy for viral particles both tested negative on faecal sampling. The owners declined treatment and the puppy was euthanased. The postmortem revealed enteric necrosis, purulent meningoencephalitis, necropurulent hepatitis and diffuse interstitial pneumonia, with heavy Salmonella enterica serovar Typhimurium growth on blood and tissue culture. The Salmonella species were sensitive to most commonly used antimicrobials including ampicillin. Canine parvovirus enteritis was diagnosed by positive canine parvovirus specific immune-peroxidase staining of intestinal tissue sections. To the authors’ knowledge, this is the first paper to describe canine parvoviral enteritis complicated by a salmonella bacteraemia, and the detection of a bacteraemia on a peripheral blood smear in a live dog.

scholarly journals Development and Application of an Immunohistochemical Staining Technique to Detect Avian Polyomaviral Antigen in Tissue Sections

1995 ◽  
Vol 7 (4) ◽  
pp. 444-450 ◽  
Author(s):  
Scott D. Fitzgerald ◽  
Willie M. Reed ◽  
Richard M. Fulton
Keyword(s):  
Immunohistochemical Staining ◽  
Rapid Test ◽  
Staining Technique ◽  
Intranuclear Inclusions ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Tissue Sections ◽  
Liver And Kidney ◽  
Diagnostic Samples ◽  
Avian Polyomavirus

An immunohistochemical staining technique was developed to detect polyomaviral antigens of budgerigar fledgling disease in formalin-fixed tissue sections. This technique used an indirect avidin-biotin, alkaline phosphatase labeling system with a mixture of monoclonal antibodies developed against the virus major capsid protein. The staining technique was applied retrospectively to 24 avian accessions which were originally diagnosed as budgerigar fledgling disease or avian polyomavirus infection based on microscopic findings including typical intranuclear inclusions. Immunohistochemical staining resulted in positive reactions in some tissues from 17 of 24 cases. The tissues most frequently containing typical intranuclear inclusions or positive immunohistochemical staining were the spleen, liver, and kidney. Neither of the 2 nonpsittacine cases was positive immunohistochemically. This technique may be used either as a rapid test on routinely processed diagnostic samples to confirm the presence of avian polyomavirus or for pathogenesis research studies.

scholarly journals Onset of Human Cytomegalovirus Replication in Fibroblasts Requires the Presence of an Intact Vimentin Cytoskeleton

2009 ◽  
Vol 83 (14) ◽  
pp. 7015-7028 ◽  
Author(s):  
Matthew S. Miller ◽  
Laura Hertel
Keyword(s):  
Intermediate Filaments ◽  
Human Cytomegalovirus ◽  
Viral Entry ◽  
Initial Phase ◽  
Axonal Neuropathy ◽  
Cell Tropism ◽  
Giant Axonal Neuropathy ◽  
Cmv Infection ◽  
Viral Particles

ABSTRACT Like all viruses, herpesviruses extensively interact with the host cytoskeleton during entry. While microtubules and microfilaments appear to facilitate viral capsid transport toward the nucleus, evidence for a role of intermediate filaments in herpesvirus entry is lacking. Here, we examined the function of vimentin intermediate filaments in fibroblasts during the initial phase of infection of two genotypically distinct strains of human cytomegalovirus (CMV), one with narrow (AD169) and one with broad (TB40/E) cell tropism. Chemical disruption of the vimentin network with acrylamide, intermediate filament bundling in cells from a patient with giant axonal neuropathy, and absence of vimentin in fibroblasts from vimentin−/− mice severely reduced entry of either strain. In vimentin null cells, viral particles remained in the cytoplasm longer than in vimentin+/+ cells. TB40/E infection was consistently slower than that of AD169 and was more negatively affected by the disruption or absence of vimentin. These findings demonstrate that an intact vimentin network is required for CMV infection onset, that intermediate filaments may function during viral entry to facilitate capsid trafficking and/or docking to the nuclear envelope, and that maintenance of a broader cell tropism is associated with a higher degree of dependence on the vimentin cytoskeleton.

Pancreatic Necrosis and Ventricular Erosion in Adenovirus-associated Hydropericardium Syndrome of Broilers

2002 ◽  
Vol 39 (3) ◽  
pp. 403-406 ◽  
Author(s):  
K. Nakamura ◽  
H. Tanaka ◽  
M. Mase ◽  
T. Imada ◽  
M. Yamada
Keyword(s):  
Broiler Chickens ◽  
Pancreatic Necrosis ◽  
Hepatic Necrosis ◽  
Pancreatic Acinar Cells ◽  
Intranuclear Inclusions ◽  
Avian Adenovirus ◽  
Hydropericardium Syndrome ◽  
Group I ◽  
Viral Particles ◽  
Broiler Farm

Seven 19-day-old broiler chickens affected with hydropericardium syndrome (HPS) with pancreatic necrosis and gizzard erosions were investigated pathologically and virologically. Mortality increased after 13 days of age in a flock on a broiler farm. The mortality rate of the flock reached 10% by 19 days of age. Macroscopically, the chickens had hydropericardium (the characteristic gross change of HPS), pinpoint white foci in the pancreas, and ventricular erosions. Histologically, the chickens had multifocal hepatic necrosis with intranuclear inclusions in hepatocytes, a marked increase of macrophages in the spleen and lung, mild epicardial edema, multifocal necrosis of pancreatic acinar cells with intranuclear inclusions, focal necrosis of the ventricular koilin layer, and degeneration of the ventricular glandular epithelium with intranuclear inclusions. Immunohistochemically, intranuclear inclusions in the liver, pancreas, and ventriculi were stained positively against group I avian adenovirus (GIAAV) antigens. Ultrastructurally, 67-nm diameter viral particles were present in intranuclear inclusions. Virologically, serotype 4 of GIAAV was isolated from the liver, heart, and kidney of affected chickens. The pathologic changes of the present cases differ from previous cases of HPS; therefore, the present strain of GIAAV may have different pathogenicity for chickens than the previous virus strain of HPS.

scholarly journals Immunoelectron microscopic localization of estrogen receptor with monoclonal estrophilin antibodies.

1985 ◽  
Vol 33 (9) ◽  
pp. 915-924 ◽  
Author(s):  
M F Press ◽  
N A Nousek-Goebl ◽  
G L Greene
Keyword(s):  
Estrogen Receptor ◽  
Ultrastructural Localization ◽  
Microscopic Level ◽  
Electron Microscopic Level ◽  
Nuclear Staining ◽  
Cytoplasmic Localization ◽  
Electron Microscopic ◽  
Tissue Sections ◽  
Receptor Antibodies ◽  
Microscopic Techniques

The recent production of a series of monoclonal estrophilin (estrogen receptor) antibodies recognizing estrogen receptor derived from a wide variety of animals and target tissues permits the development of immunoelectron microscopic techniques for identifying estrogen receptor. We have determined suitable conditions for the ultrastructural localization of estrogen receptor in tissue sections. Localization of receptor was observed in the euchromatin, but not in the marginated heterochromatin or nucleoli of epithelial and stromal nuclei of human endometrium. Competition studies indicate that only estrogen receptor specifically inhibits nuclear staining. The absence of any specific cytoplasmic localization at the electron-microscopic level is consistent with earlier light-microscopic observations and suggests that the majority of the cellular pool of estrophilin exists in the nucleus of hormone-responsive cells.

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