scholarly journals Immunomycology: rapid and specific immunocytochemical identification of fungi in formalin-fixed, paraffin-embedded material.

1993 ◽  
Vol 41 (8) ◽  
pp. 1217-1221 ◽  
Author(s):  
J A Reed ◽  
B A Hemann ◽  
J L Alexander ◽  
D J Brigati
Keyword(s):  
Complement Fixation ◽  
Fungal Infections ◽  
Periodic Acid ◽  
Cross Reactivity ◽  
Periodic Acid Schiff ◽  
Formalin Fixed Paraffin ◽  
Identification Of Fungi ◽  
Formalin Fixed Paraffin Embedded ◽  
Methenamine Silver ◽  
Formalin Fixed

We report the rapid (less than 1 hr), immunocytochemical identification of various fungi in formalin-fixed, paraffin-embedded tissues using antisera originally developed for use in immunodiffusion assays. Primary antisera directed towards fungal genera including Aspergillus, Blastomyces, Candida, Coccidioides, Cryptococcus, Histoplasma, and Sporothrix were examined. The specificity of each antiserum was evaluated by the presence or absence of crossreactivity with other morphologically similar fungi in both paraffin-embedded pure fungal cultures and tissues with culture-confirmed fungal infections. Each antiserum reacted strongly with the fungus to which it had been raised, whether examined in pure culture or infected tissues. The antisera raised against Candida, Cryptococcus, and Sporothrix did not exhibit cross-reactivity with any other fungus tested. However, the antisera raised to Aspergillus, Blastomyces, Coccidioides, and Histoplasma demonstrated significant crossreactivity with other genera of fungi, thus precluding their routine use in diagnostic immunocytochemistry. The results indicate that immunocytochemistry may provide an important adjunct to other methods, such as immunodiffusion or complement fixation assays and histochemical stains such as the Grocott methenamine silver or periodic acid-Schiff, when attempts are made to specifically identify certain fungi in formalin-fixed, paraffin-embedded tissues before mycology culture results are available.

scholarly journals Immunohistochemical Detection of Melanoma-associated Antigens on Formalin-fixed, Paraffin-embedded Canine Tumors

1994 ◽  
Vol 31 (4) ◽  
pp. 455-461 ◽  
Author(s):  
A. J. Berrington ◽  
K. Jimbow ◽  
D. M. Haines
Keyword(s):  
Monoclonal Antibodies ◽  
Periodic Acid ◽  
Specific Antigen ◽  
Periodic Acid Schiff ◽  
Cytoplasmic Staining ◽  
Normal Tissues ◽  
Melanocytic Tumors ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

Seven monoclonal antibodies (Moabs), recognizing melanoma-associated antigens in human tissues, were evaluated for their ability to immunohistochemically stain formalin-fixed, paraffin-embedded canine melanomas. Only 2 Moabs, designated human melanosome-specific antigen (HMSA)-1 and HMSA-5, stained routinely processed canine melanomas, staining 21/35 (60%) and 24/35 (69%), respectively. Twenty-nine of 35 (83%) melanomas tested were stained if results of the 2 Moabs were combined. Monoclonal antibody HMSA-1 also stained neoplastic cells of 10/35 (29%) tumors of nonmelanocytic origin and some neurons and salivary gland epithelial cells in normal canine tissues. However, Moab HMSA-1 staining in the nonmelanocytic tumors, consisting of small, discrete periodic acid-Schiff-positivc cytoplasmic droplets, was readily distinguishable from the diffusely granular, cytoplasmic staining of melanocytic tumors. In addition to melanomas, Moab HMSA-5 stained melanocytes and some melanin-containing tumor cells of a pigmented basal cell tumor and melanocytes in normal canine skin. Monoclonal antibodies HMSA-1 and HMSA-5 immunohistochemically identified the majority of canine melanomas, had limited and distinguishable staining in normal tissues and nonmelanocytic tumors, and therefore may be a useful adjunct for the diagnosis of canine melanoma in formalin-fixed, paraffin-embedded tissues.

scholarly journals THE USE OF BASIC POLYSACCHARIDES IN HISTOCHEMISTRY AND CYTOCHEMISTRY: I. THE DEMONSTRATION OF ACIDIC POLYSACCHARIDES IN TISSUE SECTIONS WITH FLUORESCEIN-LABELLED ASPERGILLUS POLYSACCHARIDE

1962 ◽  
Vol 10 (1) ◽  
pp. 8-13 ◽  
Author(s):  
E. EDWARD EVANS ◽  
SIDNEY P. KENT
Keyword(s):  
Aspergillus Parasiticus ◽  
Periodic Acid ◽  
Biological Cells ◽  
Polyvalent Cation ◽  
Periodic Acid Schiff ◽  
Tissue Sections ◽  
Formalin Fixed Paraffin ◽  
Acidic Polysaccharides ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

A procedure was described for demonstrating acidic polysaccharides in formalin-fixed paraffin-embedded tissue sections using fluorescein-labelled Aspergillus polysaccharide. This material is a basic polysaccharide composed of units of galactosamine and N-acetyl galactosamine and is synthesized by the mold Aspergillus parasiticus. The distribution of acidic polysaccharides shown by the fluorescein-labelled Aspergillus polysaccharide method was similar to results with the alcian blue-periodic acid Schiff sequence and to those with fluorescein labelled deacetylated chitin. Both epithelial and connective tissue mucins were demonstrated by the fluorescein-labelled basic polysaccharide. Aspergillus polysaccharide may be considered a colorless polyvalent cation that acts as a carrier for the fluorochrome in this procedure. Since it can react with many types of biological cells as well as with various substances isolated from cells, it may have other important applications in histochemistry and cytochemistry.

scholarly journals Standardization and Validation of an Immunoperoxidase Assay for the Detection of African Horse Sickness Virus in Formalin-Fixed, Paraffin-Embedded Tissues

2009 ◽  
Vol 21 (5) ◽  
pp. 655-667 ◽  
Author(s):  
Sarah J. Clift ◽  
Mark C. Williams ◽  
Truuske Gerdes ◽  
Marie M. E. Smit
Keyword(s):  
Detection System ◽  
Cross Reactivity ◽  
African Horse Sickness ◽  
African Horse Sickness Virus ◽  
Standard Procedures ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Complex Detection ◽  
Immunoperoxidase Assay ◽  
Formalin Fixed

An immunoperoxidase assay for the detection of African horse sickness virus (AHSV) in formalin-fixed tissues is a valuable tool in the study of the pathogenesis of the disease, as well as a useful addition to existing diagnostic tests when only preserved tissues are available. An assay that uses Hamblin antiserum in a basic avidin-biotin complex detection system was standardized and validated in accordance with the guidelines of the American Association of Veterinary Laboratory Diagnosticians Subcommittee on Standardization of Immunohistochemistry. Using 128 positive cases of African horse sickness confirmed by viral isolation and serotyping and 119 negative cases from countries where the disease has never occurred, diagnostic sensitivity and diagnostic specificity were 100% in the prime target tissues of heart and lung. There was no variation in the ability of the assay to detect all 9 serotypes of AHSV, and there was no cross-reactivity with other orbiviruses in formalin-fixed tissues. The only cross-reactivity observed was in the lungs of 2 negative cases infected with Rhodococcus equi. The assay gave good results on tissues that had been fixed in formalin for up to 365 days. Nonspecific staining was minimal provided that the standard procedures for processing and staining tissues were followed. Good immunohistochemical results were also obtained on samples fixed as long as 24 hr after death. The assay, therefore, provides a robust diagnostic tool for detection of AHSV in formalin-fixed tissues, provided the analysis is done by an experienced pathologist.

scholarly journals Underestimated Amoebic Appendicitis among HIV-1-Infected Individuals in Japan

2016 ◽  
Vol 55 (1) ◽  
pp. 313-320 ◽  
Author(s):  
Taiichiro Kobayashi ◽  
Koji Watanabe ◽  
Hideaki Yano ◽  
Yukinori Murata ◽  
Toru Igari ◽  
...  
Keyword(s):  
Acute Appendicitis ◽  
Clinical Features ◽  
Periodic Acid ◽  
Periodic Acid Schiff ◽  
Content Type ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Life Threatening ◽  
Pas Staining ◽  
Hiv 1

ABSTRACT Entamoeba histolytica is not a common causative agent of acute appendicitis. However, amoebic appendicitis can sometimes be severe and life threatening, mainly due to a lack of awareness. Also, its frequency, clinical features, and pathogenesis remain unclear. The study subjects were HIV-1-infected individuals who presented with acute appendicitis and later underwent appendectomy at our hospital between 1996 and 2014. Formalin-fixed paraffin-embedded preserved appendix specimens were reexamined by periodic acid-Schiff (PAS) staining and PCR to identify undiagnosed amoebic appendicitis. Appendectomies were performed in 57 patients with acute appendicitis. The seroprevalence of E. histolytica was 33% (14/43) from the available stored sera. Based on the medical records, only 3 cases were clinically diagnosed as amoebic appendicitis, including 2 diagnosed at the time of appendectomy and 1 case diagnosed by rereview of the appendix after the development of postoperative complications. Retrospective analyses using PAS staining and PCR identified 3 and 3 more cases, respectively. Thus, E. histolytica infection was confirmed in 9 cases (15.8%) in the present study. Apart from a significantly higher leukocyte count in E. histolytica -positive patients than in negative patients (median, 13,760 versus 10,385 cells/μl, respectively, P = 0.02), there were no other differences in the clinical features of the PCR-positive and -negative groups. In conclusion, E. histolytica infection was confirmed in 9 (15.8%) of the appendicitis cases. However, only 3, including one diagnosed after intestinal perforation, were diagnosed before the present analyses. These results strongly suggest there is frequently a failure to detect trophozoites in routine examination, resulting in an underestimation of the incidence of amoebic appendicitis.

scholarly journals Degradation of fungal DNA in formalin-fixed paraffin-embedded sinus fungal balls hampers reliable sequence-based identification of fungi

2011 ◽  
Vol 49 (3) ◽  
pp. 329-332 ◽  
Author(s):  
Odile Cabaret ◽  
Guillaume Toussain ◽  
Nassera Abermil ◽  
Issam Abd Alsamad ◽  
Françoise Botterel ◽  
...  
Keyword(s):  
Formalin Fixed Paraffin ◽  
Identification Of Fungi ◽  
Formalin Fixed Paraffin Embedded ◽  
Fungal Dna ◽  
Formalin Fixed

scholarly journals C4d immunohistochemistry in membranous nephropathy

2014 ◽  
Vol 6 (02) ◽  
pp. 076-079 ◽  
Author(s):  
Monalisa Hui ◽  
Megha S Uppin ◽  
Aruna K Prayaga ◽  
Sree Bhushan Raju ◽  
Liza Rajasekhar
Keyword(s):  
Basement Membrane ◽  
Membranous Nephropathy ◽  
Periodic Acid ◽  
Polyclonal Antiserum ◽  
Hepatitis C Infection ◽  
Periodic Acid Schiff ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Fresh Frozen ◽  
Hematoxylin And Eosin

ABSTRACT Background: Membranous nephropathy (MN) is the most common cause of nephropathy in adults. The diagnosis is based on characteristic light microscopic, electron microscope and immunofluorescence (IF) findings. In early MN, the light microscopic findings may be difficult to differentiate from minimal chain disease. In the absence of fresh frozen tissue for IF, immunohistochemistry with C4d aids in the diagnosis. Materials and Methods: A total 48 cases of MN diagnosed on renal biopsy were analyzed. The formalin fixed paraffin embedded tissues were stained with routine hematoxylin and eosin stains along with periodic acid-Schiff and silver methenamine stains to highlight the basement membrane. Fresh frozen tissues were available for IF in 40 cases. Immunostaining with C4d was done on paraffin-embedded sections by polymer-Horse Radish Peroxidase (HRP) technique using polyclonal antiserum to C4d (Biogenex, India). Results: There were 25 cases of idiopathic MN, 17 cases of Class V lupus nephritis and 2 cases were secondary to hepatitis C infection with cirrhosis. The glomerular basement membrane (GBM) was diffusely thickened with formation of spikes in 28 cases. In 11 cases the capillary loops were rigid but spikes were not seen and in 9 cases there was no apparent thickening of the basement membrane. All the cases showed diffuse positivity for C4d along the GBM. Conclusion: C4d is a reliable method to establish the diagnosis of MN and also a sensitive marker of complement activation reflecting the pathogenesis of MN.

Histopathology of fungal disease

2017 ◽  
Author(s):  
Sebastian B. Lucas
Keyword(s):  
Fungal Infections ◽  
Critical Role ◽  
Species Level ◽  
Molecular Diagnostic ◽  
Species Discrimination ◽  
Autopsy Material ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed ◽  
Made In

Histopathology has a critical role in the diagnosis of fungal infections. Often it is the first or only sample of a lesion. A rapid, confident diagnosis can significantly affect patient management. However, the morphologies of yeast and hyphae are not necessarily diagnostic at the genus or species level, and the experience of histopathologists is variable. A primary decision is whether the lesion is fungal or another infection or not infectious at all, and the next is whether the fungus is a yeast or a hyphal (mould) infection. Further histopathological genus and species discrimination can be made in many cases, but not all. Increasingly, molecular diagnostic DNA technology works effectively on formalin-fixed paraffin-embedded biopsy/autopsy material, and such information can be added to the multidisciplinary input for an optimal diagnosis.

Diagnostic value of fluorescein-labeled chitinase staining in formalin-fixed and paraffin-embedded tissues of fungal disease

2019 ◽  
Vol 58 (1) ◽  
pp. 66-70 ◽  
Author(s):  
Jin Shao ◽  
Yinggai Song ◽  
Yabin Zhou ◽  
Zhe Wan ◽  
Ruoyu Li ◽  
...  
Keyword(s):  
Fungal Infections ◽  
Periodic Acid ◽  
Poor Performance ◽  
High Sensitivity ◽  
Diagnostic Value ◽  
Fungal Disease ◽  
Periodic Acid Schiff ◽  
Ffpe Tissues ◽  
Tissue Specimens ◽  
Formalin Fixed

Abstract Common histopathologic techniques are used to diagnose fungal infections, but the diagnostic identification of mycoses in tissue specimens is often difficult, particularly when fungi rarely occur in a specimen. The aim of this study was to evaluate the application of fluorescein-labeled chitinase staining to formalin-fixed and paraffin-embedded (FFPE) tissues. We studied 79 archival FFPE tissues from patients diagnosed with fungal disease, including 38 cases of sporotrichosis and 41 cases of other fungal infections. The tissue sections were subjected to periodic acid-Schiff (PAS) staining, Gomori's methenamine silver (GMS) staining, and fluorescein-labeled chitinase staining to detect fungal elements. Culture- and/or hematoxylin-eosin-positive samples were used to estimate the diagnostic sensitivity of each staining method, with the results showing that PAS, GMS, and fluorescein-labeled chitinase staining had sensitivities of 50.6, 70.9, and 68.4%, respectively. The three staining results were the same for all fungal infections except for sporotrichosis and chromoblastomycosis. Fluorescein-labeled chitinase staining exhibited high sensitivity in cases of sporotrichosis and poor performance in detecting muriform cells of chromoblastomycosis. On the whole, the sensitivity of fluorescein-labeled chitinase staining was greater than that of PAS and similar to that of GMS staining. Therefore, the results of our study suggest that fluorescein-labeled chitinase staining is a potentially useful diagnostic tool in the diagnosis of fungal infections.

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