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.

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 Necrotizing Encephalitis Caused by Disseminated Aspergillus Infection after Orthotopic Liver Transplantation

2015 ◽  
Vol 9 (1) ◽  
pp. 1-6 ◽  
Author(s):  
Luis E. Barrera-Herrera ◽  
Alonso Vera ◽  
Johanna Álvarez ◽  
Rocio Lopez
Keyword(s):  
Liver Transplantation ◽  
Orthotopic Liver Transplantation ◽  
Fungal Infections ◽  
Periodic Acid ◽  
Surgical Techniques ◽  
Mortality Rates ◽  
Variceal Hemorrhage ◽  
Early Recovery ◽  
Periodic Acid Schiff ◽  
Aspergillus Infection

Liver transplantation is the only available treatment for some patients with end-stage liver disease. Despite reduction in mortality rates due to advances related to surgical techniques, intensive medical management and immunosuppressive therapy, invasive fungal infections remain a serious complication in orthotopic liver transplantation. We report the case of an 18-year-old male diagnosed with autoimmune cirrhosis in 2009 who was assessed and listed for liver transplantation for massive variceal hemorrhage. One year after listing a successful orthotopic liver transplantation was performed. Uneventful early recovery was achieved; however, he developed pulmonary and neurological Aspergillus infection 23 and 40 days after surgery, respectively. Antibiotic therapy with voriconazole and amphotericin was started early, with no major response. Neuroimaging revealed multiple right frontal and right parietal lesions with perilesional edema; surgical management of the brain abscesses was performed. A biopsy with periodic acid-Schiff and Gomori stains revealed areas with mycotic microorganisms morphologically consistent with Aspergillus, later confirmed by culture. The patient developed necrotizing encephalitis secondary to aspergillosis and died. Necrotizing encephalitis as a clinical presentation of Aspergillus infection in an orthotopic liver transplant is not common, and even with adequate management, early diagnosis and prompt antifungal treatment, mortality rates remain high.

scholarly journals Development and Evaluation of Nucleic Acid-Based Techniques for an Auxiliary Diagnosis of Invasive Fungal Infections in Formalin-Fixed and Paraffin-Embedded (FFPE) Tissues

2012 ◽  
Vol 53 (4) ◽  
pp. 241-245 ◽  
Author(s):  
Minoru Shinozaki ◽  
Yoichiro Okubo ◽  
Daisuke Sasai ◽  
Haruo Nakayama ◽  
Takao Ishiwatari ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Fungal Infections ◽  
Invasive Fungal Infections ◽  
Ffpe Tissues ◽  
Formalin Fixed

PodoSighter: A Cloud-Based Tool for Label-Free Podocyte Detection in Kidney Whole Slide Images

2021 ◽  
pp. ASN.2021050630
Author(s):  
Darshana Govind ◽  
Jan Becker ◽  
Jeffrey Miecznikowski ◽  
Avi Rosenberg ◽  
Julien Dang ◽  
...  
Keyword(s):  
Web Application ◽  
Periodic Acid ◽  
High Sensitivity ◽  
Ground Truth ◽  
Renal Diseases ◽  
Label Free ◽  
Current Standard ◽  
Periodic Acid Schiff ◽  
Glomerular Diseases ◽  
Whole Slide Images

Background: Podocyte depletion precedes progressive glomerular damage in several renal diseases. However, the current standard of visual detection and quantification of podocyte nuclei from brightfield microscopy images is laborious and imprecise. Methods: We have developed PodoSighter, an online cloud-based tool, to automatically identify and quantify podocyte nuclei from giga-pixel brightfield whole-slide images (WSIs) using deep learning. Ground-truth to train the tool used immunohistochemically or immunofluorescence-labeled images from a multi-institutional cohort of 122 histologic sections from mouse, rat, and human kidneys. To demonstrate generalizability of our tool in investigating podocyte loss in clinically relevant samples, we tested it in rodent models of glomerular diseases, including diabetic kidney disease, crescentic glomerulonephritis, and dose-dependent direct podocyte toxicity and depletion, as well as in human biopsies from steroid resistant nephrotic syndrome and from human autopsy tissues. Results: The optimal model yielded high sensitivity/specificity of 0.80/0.80, 0.81/0.86, and 0.80/0.91, in mouse, rat, and human images, respectively, from periodic-acid Schiff-stained WSIs. Furthermore, the podocyte nuclear morphometrics extracted using PodoSighter were informative in identifying diseased glomeruli. We have made PodoSighter freely available to the general public as turnkey plugins in a cloud-based web application for end-users. Conclusion: Our study demonstrates an automated computational approach to detect and quantify podocyte nuclei in standard histologically-stained WSIs, facilitating podocyte research and enabling possible future clinical applications.

Morphological evaluation of canine platelets on Giemsa- and PAS-stained blood smears

2005 ◽  
Vol 53 (3) ◽  
pp. 337-350 ◽  
Author(s):  
Dóra Halmay ◽  
P. Sótonyi ◽  
P. Vajdovich ◽  
T. Gaál
Keyword(s):  
Diabetes Mellitus ◽  
Skin Diseases ◽  
Periodic Acid ◽  
Morphological Characteristics ◽  
Diagnostic Value ◽  
Endocrine Disorders ◽  
Periodic Acid Schiff ◽  
Giant Platelets ◽  
Immune Mediated ◽  
Blood Smears

The morphology of canine platelets (changes in size, shape, staining characteristics, degree of activation and clump formation, distribution of granules, appearance of vacuoles on Giemsa-stained smears) was investigated in 20 healthy control and 181 diseased dogs. In the group of the sick dogs 84 animals suffered from disorders affecting directly the haematological parameters or the haematopoietic organs such as bleeding, thymic haemorrhage, haemolytic disorders, lymphoma, immune-mediated thrombocytopenia, and other 97 dogs were affected by other diseases (hepatopathy, nephropathy, hepatic, splenic or intestinal neoplasm, skin diseases, diabetes mellitus, Cushing'ssyndrome, sepsis). The alterations found in platelet morphology were not specific for any disorder. The most common platelet abnormalities were polychromasia and the presence of giant platelets. These changes occurred in a high number in disorders accompanied by bleeding or haemolysis. Anisocytosis was the most frequent finding in hepatic, splenic or intestinal neoplasms and in certain endocrinopathies. Microcytosis was observed in immune-mediated thrombocytopenia, hepatic neoplasms and endocrine disorders. Extreme platelet activation was common in haemolysis, hepatopathies, neoplastic diseases and sepsis. Vacuolisation was present in thymic haemorrhage, pancreatitis, diabetes mellitus and Cushing's syndrome. A new morphologic phenomenon, i.e. a ring-like formation of granules, was described in the cytoplasm of the platelets both in healthy and diseased animals. In addition, two forms of pathologic granulation were also described for the first time in Giemsa-stained blood smears: the pseudonuclear and the spot-like formation of granules, which were observed especially in disorders affecting the blood cells. The granulation and morphological characteristics of platelets on smears stained by periodic acid-Schiff reaction (PAS) were also studied. Three localisations of granulation were observed, such as peripheral, eccentric and diffuse. The ratio of PAS-positive and -negative platelets was evaluated in several diseases. Our findings support the diagnostic value of platelet evaluation by light microscopy and help clinicians/clinical pathologists to understand why morphologic changes of thrombocytes might be expected in several diseases.

scholarly journals Technical Aspects and Applications for Developing in situ Hybridization Procedures for Formalin-Fixed and Paraffin-Embedded (FFPE) Tissues for Diagnosis of Fungal Infections

2017 ◽  
Vol 58 (1) ◽  
pp. E33-E37 ◽  
Author(s):  
Minoru Shinozaki ◽  
Naobumi Tochigi ◽  
Sota Sadamoto ◽  
Somay Yamagata Murayama ◽  
Megumi Wakayama ◽  
...  
Keyword(s):  
Fungal Infections ◽  
Technical Aspects ◽  
Ffpe Tissues ◽  
Formalin Fixed

Role of FNA and Special Stains in Rapid Cytopathological Diagnosis of Pulmonary Nocardiosis

2018 ◽  
Vol 62 (3) ◽  
pp. 178-182 ◽  
Author(s):  
Ridhi Sood ◽  
Ruchita Tyagi ◽  
Pavneet Kaur Selhi ◽  
Gursheen Kaur ◽  
Harpreet Kaur ◽  
...  
Keyword(s):  
Fungal Infections ◽  
Periodic Acid ◽  
Tertiary Care ◽  
Opportunistic Pathogen ◽  
Clinical Findings ◽  
Accurate Diagnosis ◽  
Care Hospital ◽  
Periodic Acid Schiff ◽  
Pulmonary Nocardiosis ◽  
Inflammatory Infiltrates

Background: Nocardia, a gram-positive aerobic bacillus of the Actinomycetales family, is a significant opportunistic pathogen in immunocompromised individuals. Clinical and radiological features of pulmonary nocardiosis are nonspecific and can be misdiagnosed as tuberculosis, pneumocystis, staphylococcal or fungal infections, or as malignancy. Aspiration cytology with special stains is a quick and effective approach for accurate diagnosis. Materials and Methods: We present 7 cases of pulmonary nocardiosis, admitted to the pathology department in a tertiary-care hospital in Punjab. Clinical findings, immune status, laboratory tests, chest radiographs, and computed tomography scans were reviewed. Cytologically, special stains like 1% Ziehl-Neelsen (ZN), 20% ZN, periodic acid-Schiff (PAS), Grocott methenamine silver (GMS), and reticulin stains were studied along with May-Grünwald Giemsa, Papanicolaou, and hematoxylin and eosin. Results: All the patients were immunocompromised. The radiological changes were nonspecific. Cytomorphology showed acute and chronic inflammatory infiltrates with necrosis. None of the cases showed well-defined granulomas. GMS, modified 1% ZN and, Gordon and Sweet reticulin stains highlighted the delicate filamentous bacteria in all cases. PAS and 20% ZN stain for tuberculous bacilli were uniformly negative. Conclusion: FNAC can provide a quick and accurate diagnosis of nocardiosis and thereby facilitate timely medical management.

scholarly journals THE USE OF BASIC POLYSACCHARIDES IN HISTOCHEMISTRY AND CYTOCHEMISTRY; II. THE DEMONSTRATION OF ACIDIC POLYSACCHARIDES IN TISSUE SECTIONS USING FLUORESCEIN-LABELLED DEACETYLATED CHITIN

1962 ◽  
Vol 10 (1) ◽  
pp. 14-18 ◽  
Author(s):  
SIDNEY P. KENT ◽  
E. EDWARD EVANS
Keyword(s):  
Connective Tissue ◽  
Acridine Orange ◽  
Alcian Blue ◽  
Periodic Acid ◽  
Fluorescent Staining ◽  
Periodic Acid Schiff ◽  
Tissue Sections ◽  
Acidic Polysaccharides ◽  
Staining Methods ◽  
Formalin Fixed

A method for demonstrating acidic polysaccharides in formalin fixed parafin embedded tissue sections using fluorescein labelled deacetylated chitin is described. Epithelial and connective tissue acidic polysaccharides have been studied in a number of organs. The distribution of acidic polysaccharides as seen with fluorescein-labelled deacetylated chitin is similar to results obtained with alcian blue-periodic acid Schiff and fluorescein-labelled Aspergillus polysaccharide. Previously reported fluorescent staining methods for acidic polysaccharides such as iron-hematoxylin-acridine orange and atabrine as well as fluorescein-labelled deacetylated chitin may prove to be useful adjuncts to the light microscopic methods of demonstrating mucins. Other possible uses of deacetylated chitin, a colorless macro-cation, in histochemistry and cytochemistry are noted.

scholarly journals Evaluation of mucin histochemistry in benign and malignant prostatic lesion and their correlationPSA level

2014 ◽  
Vol 4 (8) ◽  
pp. 612-616
Author(s):  
S Bastola ◽  
OP Talwar
Keyword(s):  
Alcian Blue ◽  
Statistical Significance ◽  
Periodic Acid ◽  
Malignant Lesion ◽  
Diagnostic Value ◽  
Chi Square ◽  
Periodic Acid Schiff ◽  
Mucin Histochemistry ◽  
Psa Density ◽  
Chi Square Test

Background: The major causes of prostatic enlargement are nodular hyperplasia, prostatitis and neoplasm including prostatic intraepithelial neoplasm. Acidic mucin is absent in benign prostatic glands and is present in some prostatic adenocarcinomas. Mucin stain may be an adjunctive aid in the diagnosis of prostatic adenocarcinoma. This study aims to find out role of acidic mucin in differentiating benign and malignant lesion together with the aid of PSA level.Materials and Methods: A study was done from August 2006 to August 2008. Serum PSA level was performed. Biopsy specimen were processed and stained with Hematoxylin and Eosin as well as Alcian Blue and Periodic Acid Schiff. Data was coded and entered into SPSS v. 11 and Chi-square test and scatter plot was done to analyze the findings.Results: Out of 207 prostatic specimens studied, 78.3% showed BPH with or without prostatitis whereas 12% showed PIN and 8.7% malignancy. PSA density showed statistical significance with p= 0.02, in favor of malignancy. Alcian blue positivity was in 77.8% of malignant cases and 18.5 % in benign cases (p = 0.000003). Acid mucin was more in well differentiating tumours decreasing significantly in high grade malignancies. Combined diagnostic value of PSA>10 ng/ml and Alcian blue positivity showed increase in specificity to 96.8% for diagnosing a malignant lesion, with positive predictive value of 57.1%.Conclusion: PSA density can be used to identify and pick up malignant cases where intermediate levels of PSA values are common. Alcian blue stain can help in assessing the degree of differentiation in malignant cases. DOI: http://dx.doi.org/10.3126/jpn.v4i8.11611 Journal of Pathology of Nepal; Vol.4,No8(2014) 612-616       

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.

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