scholarly journals Constant Detection of CD2, CD3, CD4, and CD5 in Fixed and Paraffin-embedded Tissue Using the Peroxidase-mediated Deposition of Biotin-Tyramide

1997 ◽  
Vol 45 (12) ◽  
pp. 1665-1672 ◽  
Author(s):  
Rainer Malisius ◽  
Hartmut Merz ◽  
Boris Heinz ◽  
Evariste Gafumbegete ◽  
Britta U. Koch ◽  
...  
Keyword(s):  
Binding Sites ◽  
Lymphoid Tissues ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Leukocyte Antigens ◽  
Formalin Fixed Paraffin Embedded ◽  
Tissue Specimens ◽  
Immunohistochemical Techniques ◽  
Microwave Techniques ◽  
Formalin Fixed

Immunohistochemical methods are widely used for diagnostic purposes in histopathology. However, the use of most monoclonal anti-leukocyte antibodies is limited to frozen tissues. Initially, it was believed that formalin fixation in particular, which is the gold standard for morphological tissue preservation, destroys most of the antigen binding sites. In recent years, protease digestion and the introduction of microwave techniques have significantly enhanced the sensitivity of immunohistochemical techniques, and a variety of hidden antigen sites in formalin-fixed tissue have been retrieved for initially unreactive antibodies. It therefore became clear that many of the leukocyte antigens are not irreversibly destroyed but are most probably masked during the fixation process. We developed a technique combining optimized pretreatment of formalin-fixed tissue with a dramatic enhancement of the immunohistochemical sensitivity and named it the ImmunoMax method. The ImmunoMax method proves that by optimizing the technique at the following three levels it is possible to detect formalin-sensitive leukocyte antigens: (a) standard fixation of the tissue; (b) sufficient antigen unmasking; and (c) increasing the substrate turnover by multiplication of binding sites with subsequent enhancement of the immunohistochemical reaction. Using this optimized ImmunoMax method, we were able to detect CD2, CD3, CD4, and CD5 with conventional monoclonal antibodies in formalin-fixed, paraffin-embedded tissue specimens of various lymphoid tissues.

scholarly journals Antigen retrieval in formalin-fixed, paraffin-embedded tissues: an enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections.

1991 ◽  
Vol 39 (6) ◽  
pp. 741-748 ◽  
Author(s):  
S R Shi ◽  
M E Key ◽  
K L Kalra
Keyword(s):  
Polyclonal Antibodies ◽  
Antigen Retrieval ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Tissue Sections ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Pre Treatment ◽  
Immunohistochemical Techniques ◽  
Formalin Fixed

We describe a new approach for retrieval of antigens from formalin-fixed, paraffin-embedded tissues and their subsequent staining by immunohistochemical techniques. This method of antigen retrieval is based on microwave heating of tissue sections attached to microscope slides to temperatures up to 100 degrees C in the presence of metal solutions. Among 52 monoclonal and polyclonal antibodies tested by this method, 39 antibodies demonstrated a significant increase in immunostaining, nine antibodies showed no change, and four antibodies showed reduced immunostaining. In particular, excellent immunostaining results were obtained with a monoclonal antibody to vimentin as well as several different keratin antibodies on routine formalin-fixed tissue sections after pre-treatment of the slides with this method. These results showed that after antigen retrieval: (a) enzyme predigestion of tissues could be omitted; (b) incubation times of primary antibodies could be significantly reduced, or dilutions of primary antibodies could be increased; (c) adequate staining could be achieved in long-term formalin-fixed tissues that failed to stain by conventional methods; and (d) certain antibodies which were typically unreactive with formalin-fixed tissues gave excellent staining.

scholarly journals A simple technique for preservation of fixation-sensitive antigens in paraffin-embedded tissues.

1994 ◽  
Vol 42 (8) ◽  
pp. 1127-1134 ◽  
Author(s):  
J H Beckstead
Keyword(s):  
Simple Technique ◽  
Frozen Section ◽  
Lymphoid Tissues ◽  
Frozen Sections ◽  
Morphological Examination ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Important Approach ◽  
Reduced Toxicity ◽  
Formalin Fixed

Immunohistochemistry is a powerful tool for tissue diagnosis and research. Although the frozen section has remained the gold standard for this important approach to evaluation of antigens in tissues, there is widespread acknowledgment of many limitations. Routine paraffin-embedded sections ware widely used for morphological examination of tissues but are not optimal for antigen preservation. In this study, paraffin-embedded tissues fixed with a simple buffer containing zinc as the primary fixative were compared with tissues fixed with routine formalin, zinc-formalin, paraformaldehyde, ethanol, a variety of commercial (non-formalin-containing) fixatives that have been recommended for reduced toxicity and improved antigen survival, and frozen sections. Human lymphoid tissues and a group of antibodies to antigens (CD1, CD4, CD7, CD8, CD19) usually preserved only in frozen tissue were used as a model system. Fixation in a simple solution of zinc acetate and zinc chloride in a Tris-Ca acetate buffer resulted in antigen preservation comparable to that in frozen sections with antibodies to these cell surface markers. Morphological preservation was comparable to formalin-fixed sections. The work presents a new method that represents the closest approach yet to a technique that combines optimal antigenic survival with the convenience and morphological preservation of traditional formalin-fixed tissue embedded in paraffin.

scholarly journals 419. Diagnostic Performance of Immunohistochemistry Test to Differentiate Aspergillosis from Mucormycosis With Formalin-Fixed Tissue Specimens

2018 ◽  
Vol 5 (suppl_1) ◽  
pp. S159-S160
Author(s):  
Ji Hyun Yun ◽  
Sungim Choi ◽  
Jung Wan Park ◽  
Kyung Hwa Jung ◽  
Kyeong Min Jo ◽  
...  
Keyword(s):  
Diagnostic Performance ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Tissue Specimens ◽  
Formalin Fixed

scholarly journals An Immunohistochemical Study on the Cytogenetic Origin of Pulmonary Multinucleate Giant Cells in Porcine Dermatosis Vegetans

1993 ◽  
Vol 30 (2) ◽  
pp. 162-170 ◽  
Author(s):  
O. Evensen
Keyword(s):  
Monoclonal Antibodies ◽  
Positive Reaction ◽  
Alveolar Macrophages ◽  
Giant Cells ◽  
Immunohistochemical Method ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Tissue Specimens ◽  
Formalin Fixed ◽  
Multinucleate Giant Cells

The origin of pulmonary multinucleate giant cells (MGC) in porcine dermatosis vegetans was studied in six Norwegian Landrace pigs ages 4 (male), 5 (female), 6 (female), 10 (female), and 12 (one male, one female) weeks, using an avidin biotin peroxidase and alkaline phosphatase complex immunohistochemical method on sections of formalin- and ethanol-fixed and frozen tissue specimens. Well-characterized, commercially available antisera/monoclonal antibodies to keratin, vimentin, lysozyme, a monocytic antigen, and a myelomonocytic antigen were used. The immunoreactivity to intermediate-sized filaments in MGC was negative for keratins and positive for vimentin. In addition, a positive reaction was found in alveolar macrophages, chondrocytes, fibrocytes, alveolar lymphocytes, and granulocytes in ethanol-fixed tissue. Marked masking was observed in formalin-fixed tissue. Antilysozyme antiserum gave a positive cytoplasmic reaction in alveolar macrophages and MGC, although the reaction was variable in ethanol-fixed tissue. In trypsinized formalin-fixed tissue, a moderate and more consistent cytoplasmic reaction was observed in alveolar macrophages and MGC. Two monoclonal antibodies that identify human cells of the MMS, EMB 11 and Mac 387, were negative for EMB 11 in ethanol-fixed and frozen sections, whereas Mac 387 showed a positive and specific cytoplasmic immunoreaction in alveolar macrophages and small MGC in ethanol- and formalin-fixed tissue. Pulmonary MGC in dermatosis vegetans are derived from mesenchymal cells, and substantial evidence is provided in support of a monocyte/macrophage origin based on a positive reaction for lysozyme and a myelomonocytic antigen. The importance of adequate fixatives for immunohistochemical demonstration of cell-specific markers is clearly shown.

scholarly journals Effects of fixation time and enzymatic digestion on immunohistochemical demonstration of bromodeoxyuridine in formalin-fixed, paraffin-embedded tissue.

1988 ◽  
Vol 36 (5) ◽  
pp. 511-514 ◽  
Author(s):  
Y Hayashi ◽  
M Koike ◽  
M Matsutani ◽  
T Hoshino
Keyword(s):  
Human Brain ◽  
Enzymatic Digestion ◽  
Fixation Time ◽  
Reaction Products ◽  
Human Brain Tumor ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

We studied the effects of fixation time and enzymatic digestion on immunohistochemical staining for bromodeoxyuridine (BUdR) in excised rat and human gastrointestinal tissues and human brain tumors which had been fixed in formalin after intravenous administration of BUdR shortly before biopsy of tissue. In formalin-fixed rat gastrointestinal tissues not treated with proteinase, the reaction products were insufficient to identify BUdR-positive cells. Results similar to those in ethanol-fixed tissue were obtained when formalin-fixed tissue sections were treated with protease, pepsin, or trypsin. The longer the material had been fixed in formalin, the longer the incubation in proteinase required to identify BUdR-labeled nuclei. The BUdR labeling indices of formalin-fixed human brain tumor specimens treated with protease were comparable to those of ethanol-fixed tissues. Sufficient BUdR staining was obtained even in tissues fixed in formalin for prolonged periods. Therefore, the BUdR labeling index can be determined retrospectively in clinical materials stored in formalin.

scholarly journals Molecular Diagnosis of Bordetella pertussis Infection by Evaluation of Formalin-Fixed Tissue Specimens

2006 ◽  
Vol 44 (3) ◽  
pp. 1074-1076 ◽  
Author(s):  
K. M. Tatti ◽  
K.-H. Wu ◽  
G. N. Sanden ◽  
P. Greer ◽  
J. Sumner ◽  
...  
Keyword(s):  
Molecular Diagnosis ◽  
Bordetella Pertussis ◽  
Pertussis Infection ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Tissue Specimens ◽  
Formalin Fixed

Application of anti-BCG antibody for rapid immunohistochemical detection of bacteria, fungi and protozoa in formalin-fixed paraffin-embedded tissue samples

2008 ◽  
Vol 56 (1) ◽  
pp. 89-99 ◽  
Author(s):  
Levente Szeredi ◽  
Róbert Glávits ◽  
Miklós Tenk ◽  
Szilárd Jánosi
Keyword(s):  
Bacterial Species ◽  
Histochemical Staining ◽  
Fixed Tissue ◽  
Tissue Samples ◽  
Bacteriological Culture ◽  
Formalin Fixed Tissue ◽  
Formalin Fixed Paraffin Embedded ◽  
Almost All ◽  
Formalin Fixed

The applicability of an anti- Mycobacterium bovis (BCG) antibody-based immunohistochemistry (IHC) procedure was investigated using everyday veterinary pathological samples collected from 13 different animal species. Fifty-one formalin-fixed and paraffin-embedded tissue samples were selected for this study. Forty, 4 and 7 tissue samples contained different species of bacteria, fungi and protozoa, respectively. Three serial sections were prepared in each case. Two sections were pre-treated with enzyme and heat, respectively, while the last section was not pre-treated. In seven cases the sensitivity of histochemical staining (HSM), IHC and bacteriological culture were compared. Heating of the sections in a microwave oven was the most effective method in the case of almost all pathogens used. Strong or moderate positive reactions were observed for 26 bacterial species, all fungal and 2 protozoal species, while weak reactions occurred for 2 bacterial and 1 protozoal species. Only 4 protozoal and 12 bacterial species, including Leptospira and all the five Mycoplasma species examined, showed no reaction in this test. IHC had almost the same sensitivity as bacteriological culture and was more sensitive than HSM. The IHC method presented here should be preferred to HSM as a general screening tool in cases where pathological lesions suspicious for infections are evident and no microorganism can be cultured in vitro or only formalin-fixed tissue samples are available for the laboratory examination.

Retrospective Analysis of Ploidy in Primary Osseous and Extraosseous Ewing Family Tumors in Children

1998 ◽  
Vol 84 (4) ◽  
pp. 493-498 ◽  
Author(s):  
Daniela Perotti ◽  
Valentina Corletto ◽  
Roberto Giardini ◽  
Antonina Parafioriti ◽  
Franca Fossati-Bellani ◽  
...  
Keyword(s):  
Ewing’S Sarcoma ◽  
Ewing's Sarcoma ◽  
Image Cytometry ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Frozen Tissue ◽  
Formalin Fixed Paraffin Embedded ◽  
Fresh Frozen ◽  
Ewing Family Tumors ◽  
Formalin Fixed

Aims To restrospectively study the DNA content in a series of childhood Ewing Family Tumors (EFT), and to investigate its prognostic value. Methods The study was performed on a series of 27 EFTs (osseous Ewing's sarcoma, 18 cases; extraos-sous Ewing's sarcoma, 2; peripheral neuroepithelioma, 4; Askin Rosai tumors, 3). Ploidy was investigated using both flow cytometry (FCM) and image cytometry (ICM) on tumor cell suspensions from formalin-fixed paraffin-embedded specimens or fresh frozen tissue obtained from the primary tumor at diagnosis. Results Ploidy was evaluable by FCM in all cases, and by ICM in 23/27. When fresh frozen tissue and paraffin-embedded samples from the same tumor were available for analysis, they yielded equal results. The rate of agreement between FCM and ICM was 82%. The majority of cases were diploid, and in the present series aneuploidy seemed to be associated with a poor outcome. Conclusions These results suggest that aneuploidy could be an indicator of a bad prognosis in EFT; however, the small number of cases precludes any conclusion of statistical value. Larger restrospective studies on ploidy using archival material could be performed and their reliability is supported by the concordance of results from fresh and formalin-fixed tissue.

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