Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Tissue Embedding”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 505 records · Page 28Linked to original sources
PubMed1993

Immunohistochemical detection of swine influenza A virus in formalin-fixed and paraffin-embedded tissues.

An avidin-biotin complex (ABC) immunohistochemical method utilizing a commercially-available polyclonal antiserum to human influenza A virus was used to detect antigens of influenza A virus in formalin-fixed, paraffin-embedded tissues of swine. Influenza A antigens were immunohistochemically detected in 28/30 cases in which influenza A virus was demonstrated by virus isolation and in 5/22 cases suspected to be influenza A-infected by clinical and histological criteria, but from which the virus was not isolated. Viral antigen was not demonstrated in 30/30 cases not suspected clinically or histologically to be associated with influenza infection. This method is a convenient, sensitive, and specific means of influenza A virus detection and is applicable to both routine diagnosis of influenza A virus infection and to retrospective and prospective studies of the occurrence and the pathogenesis of this virus in pigs.

Animals↗
PubMed1999

Diagnosis of malignant catarrhal fever by PCR using formalin-fixed, paraffin-embedded tissues.

A previously described polymerase chain reaction (PCR) assay (amplification of a 238-bp fragment of ovine herpesvirus 2 [OHV-2] genomic DNA) for diagnosis of sheep-associated malignant catarrhal fever (MCF) was adapted for use on formalin-fixed, paraffin-embedded tissues. Variables affecting its use were examined. Archived tissues from cattle, white-tailed deer (Odocoileus virginianus), and bison (Bison bison) diagnosed with MCF by clinical signs or histologic lesions were obtained from 2 veterinary diagnostic laboratories. Tissues from healthy animals and from animals diagnosed with other common bovine viral diseases were examined as controls. A total of 86 blocks from 37 suspect MCF cases were examined. Forty-one blocks from healthy animals and animals with unrelated viral diseases were examined as controls. The assay was specific for sheep-associated MCF and did not yield false-positive signals from healthy animals or from cases of infectious bovine rhinotracheitis, bovine virus diarrhea, mucosal disease, or parainfluenza-3 virus infection. A wide variety of tissues were suitable substrates, including spleen, lymph node, intestine, brain, lung, and kidney. Extracted DNA provided a more suitable target than did unextracted tissue lysate. The highest levels of viral DNA were present in lymphoid organs and intestine, but the data indicate that in acute clinical cases, most organs contain sufficient viral DNA to serve as a suitable diagnostic specimen. Fixation of 0.5-cm3 blocks of tissue in 10% neutral buffered formalin was deleterious to the target DNA, and PCR signals progressively diminished after fixation for >45 days. Detection of genomic DNA of OHV-2 by PCR was successful for archived tissues that were 15 years old.

Animals↗
PubMed1997

[Flow cytometric quantification of numerical chromosome aberrations in non-small cell lung carcinomas using formalin-fixed paraffin-embedded tissue].

Fluorescence in situ hybridization with biotinylated repetitive DNA probe specific for the centromeric region of chromosome 17 (p17H8: Oncor) was applied to suspended nuclei which were isolated by Shutte's method from formalin-fixed paraffin-embedded tissue. The tissues were obtained from surgically resected specimens from nine patients with non-small cell lung carcinoma. The isolated nuclei were prepared with 0.05% pepsin/0.1NHCl for 15 minutes at 37 degrees C. Subsequently, these were immersed in 70% acetic acid for 10 seconds at room temperature. After heat denature with hybridization mixture which contained 3 mu 1 DNA probe for 10 minutesat 70 degrees C, 1 x 10(6) nuclei were incubated overnight at 37 degrees C. After washing with 60% formamide/2 x SSC, the hybridized probes were labeled by FITC conjugated avidin. A number of centromeric signals of chromosome 17 wasevaluated by fluorescence microscopy (BH-2, Olympus). Furthermore, a probe-related FITC intensity was quantified using flow cytometry (FACScan, Becton Dickinson). As the results, there was good correlation between a relative fluorescence intensity determined by flow cytometry and a relative fluorescence signal by fluorescence microscopy (p < 0.05).

Aged↗
PubMed1993

Retrospective molecular detection of Transthyretin Met 111 mutation in a Danish kindred with familial amyloid cardiomyopathy, using DNA from formalin-fixed and paraffin-embedded tissues.

Severe familial amyloid cardiomyopathy (FAC) in a Danish kindred is associated with a specific mutation (Met for Leu 111) in the transthyretin (TTR) gene. The mutation causes the loss of a DdeI restriction site in the gene, allowing molecular diagnostic studies. We studied formalin-fixed, paraffin-embedded tissues, up to 39 years old, from 29 family members of this kindred. DNA was partially purified from deparaffinized tissue sections and a DNA sequence of the TTR gene flanking the mutation site was amplified by the polymerase chain reaction (PCR), followed by restriction enzyme analysis. Amplified DNA was obtained from tissues representing 23 of the 29 persons. Ten out of the 23 family members were found to carry the TTR Met 111 mutation, whereas 13 were not affected. The results were consistent with known clinical data and with corresponding serum TTR examinations. This retrospective study shows that archival tissues can be used to confirm the diagnosis and disease pattern in members of families affected by hereditary diseases.

Adolescent↗
PubMed1999

Identification of MEN1 mutations in sporadic enteropancreatic neuroendocrine tumors by analysis of paraffin-embedded tissue.

Gastrinomas and other gastrointestinal neuroendocrine tumors may occur sporadically or as part of the inherited syndrome multiple endocrine neoplasia type 1 (MEN1). Mutations in the recently identified MEN1 gene have been described in sporadic gastrinomas and insulinomas. This study describes techniques used to identify mutations in the MEN1 gene in DNA extracted from paraffin-preserved tissue. Two novel mutations are identified in the MEN1 gene from nine archived paraffin-embedded neuroendocrine tumors, demonstrating that retrospective genetic analysis can be used to identify mutations in the MEN1 gene from preserved tissue. Conditions are provided by which paraffin-embedded tissue can be used as a source of genetic material for sequence information of sufficient quality for mutational studies of the MEN1 gene. It should also be possible to apply this retrospective genetic analysis of paraffin-embedded tissue to other disease models.

DNA, Neoplasm↗
PubMed1997

Application of fluorescence in situ hybridization to detect N-myc (MYCN) gene amplification on paraffin-embedded tissue sections of neuroblastomas.

Fluorescence in situ hybridization (FISH) was applied to neuroblastoma for detection of N-myc (MYCN) oncogene amplification, and the results were compared with Southern blot analysis (Southern). In nine neuroblastomas (formalin-fixed paraffin-embedded tissues were available in seven cases including two cases with touch preparations, and two cell lines), all five cases with N-myc amplification detected by Southern had cells with multiple N-myc signals by FISH, and three cases showed no N-myc amplification either by Southern or FISH procedure. One case, not examined by Southern, showed amplified signals of N-myc by FISH. These data indicate that FISH results for N-myc amplification have close correlation with Southern blot analysis. The chromosome 2-specific repetitive DNA probe was also applied for the analysis of ploidy by FISH. Six cases with N-myc amplification by Southern and/or FISH had diploid tumors and two cases without amplified N-myc showed aneuploidy. The remaining one case consisted of heterogeneous elements showing diploidy in undifferentiated tissue and both aneuploidy (ganglionic cells) and diploidy (Schwann cells) in differentiated area. We conclude that FISH is a practical, useful and reliable method over Southern especially for analysis of N-myc amplification in neuroblastoma, and simultaneous cohybridization with a specific chromosome probe is of great value in predicting the prognosis of patients.

Blotting, Southern↗
PubMedDate not supplied

Modifications to the in situ TUNEL method for detection of apoptosis in paraffin-embedded tissue sections.

The in situ detection of cells undergoing apoptosis is increasingly important in the analysis of injury and degeneration in the central nervous system. Limited information is presently available on the quantification of apoptosis in paraffin-embedded brain tissue sections, a technique which would be most useful in the evaluation of archival tissue for diagnostic and experimental purposes. In this report, optimized conditions for tissue digestion and permeabilization using Proteinase K and Triton X and a quantification method for apoptosis detection are described using brain sections from aluminum maltolate-treated aged and young rabbits as compared to untreated matched controls. This method provides optimal staining of apoptotic cells without the problem of tissue destruction, and should prove useful in evaluating the process of apoptosis in neurodegenerative disorders.

Aging↗
PubMed1998

Analysis of clonality in human endometriotic cysts based on evaluation of X chromosome inactivation in archival formalin-fixed, paraffin-embedded tissue.

The clonality of epithelial cells in ovarian endometriotic cysts was evaluated on the basis of a polymorphism of the X-linked phosphoglycerate kinase gene (PGK). The problems associated with clonality analysis of DNA samples extracted from archival formalin-fixed, paraffin-embedded tissue were addressed with the use of newly designed primers and a modified stepdown program for PCR amplification. The method relies on digestion of DNA with the methylation-sensitive restriction enzyme SnaBI, PCR amplification of PGK, and detection of a BstXI polymorphism. With this approach, only the inactive (methylated) PGK allele is selectively amplified by PCR. A total of 25 epithelial cells in endometriotic cysts and 25 matched normal ovarian stromal tissue specimens were analyzed. All of the control tissue samples yielded PCR products, with 11 of the 25 samples found to be heterozygous for the BstXI polymorphism. Ten of the 25 epithelial cells in endometriosis specimens were informative at this locus, and all of these 10 were shown to be monoclonal on the basis of PGK methylation. The results indicate that this new method of clonal analysis with archival formalin-fixed tissue is reliable and confirm that endometriotic cysts are monoclonal in origin.

Base Sequence↗
PubMed1996

Immunohistochemical detection of p21waf1/cip1/sdi1 and p53 proteins in formalin-fixed, paraffin-embedded tissue sections of colorectal carcinoma.

Wild-type p53 protein is a critical participant in G1 cell cycle arrest through the induction of waf1/cip1/sdi1 gene product p21, but mutant p53 proteins have lost transcriptional activity. To know whether the view obtained from studies using cultured cells is adaptable to human solid tumors, the authors immunohistochemically stained p21 and p53 in formalin-fixed, paraffin-embedded tissue sections from 67 cases of colorectal carcinoma. Of these, 54 cancers showed positive staining of p53, whereas p21-positive cells were identified in 46 tumors. However, the proportion of cells positive for these proteins varied considerably among cases, and moreover wide variation in p21 immunoreactivity was noted within the same tumor. Although there was an inverse relationship between p21 and p53 staining in tumors, namely, p21-positive cells were p53 negative and vice versa, an intermingling of tumor cells expressing concomitantly both genes was noted in 34 (51%) of 67 tumors. In these tumors, both proteins were usually moderately stained, but tumor cells intensely coexpressing both were found in one sample. This study supports the notion that although p21 expression is regulated by p53 under physiological conditions, it can be regulated by an additional pathway(s) in solid malignant tumors.

Carcinoma↗
PubMed1990

Coxsackie B antigen in the central nervous system of a patient with fatal acute encephalitis: immunohistochemical studies of formalin-fixed paraffin-embedded tissue.

A case of fatal acute encephalitis due to Coxsackie B1 virus is described. Confirmation of Coxsackie B virus as the etiological agent of encephalitis was based on identification of the virus antigen in formalin-fixed paraffin-embedded tissue sections. In the past, the diagnosis was obtained by serological studies of peripheral blood and viral isolation. This is the first report in which indirect immunofluorescent and immunoperoxidase methods using rabbit antiserum raised against Coxsackie B types 1-6 was utilized in determining the etiology of encephalitis. It must be emphasized that these methods can be used both on biopsy or autopsy specimens, even retrospectively.

Antigens, Viral↗
PubMed2002

Expression of E-cadherin in prostate cancer in formalin-fixed, paraffin-embedded tissues: correlation with pathological features.

AIMS: E-cadherin has been studied recently as a potential marker for tumour progression. The present study aimed to assess the expression of E-cadherin in formalin-fixed and paraffin-embedded radical prostatectomy specimens and to compare its expression with the pathological stage and Gleason score. METHODS: This study comprised a total of 58 men who were selected on the basis of the negative surgical margins of surgical specimens from radical retropubic prostatectomies and concomitant pelvic lymph node dissections. Indirect immunoperoxidase staining was performed as described previously using HECD-1 monoclonal antibody with the retrieval of antigen by treatment of the paraffin-embedded tissue with microwaves in citrate buffer. RESULTS: Aberrant staining patterns of E-cadherin were observed in 18 (64%) and in 25 (83%) of cases with pathological stages pT2 and pT3a, respectively (P>0.05). Immunohistochemical examination also revealed aberrant staining patterns of E-cadherin in 16 (89%) of specimens with Gleason score > or =7 and in 27 (68%) of specimens with Gleason score <7 (P>0.05). Biochemical recurrence was identified in three (5%) patients and immunohistochemical examination of their specimens revealed aberrant staining patterns of E-cadherin molecule in all of them. CONCLUSION: Our preliminary results indicate that, even though we could not demonstrate any significant correlation between E-cadherin staining pattern and tumour invasion and Gleason scores, aberrant staining patterns of E-cadherin may be a significant predictor for disease recurrence following radical prostatectomies if supported by large scale studies.

Adenocarcinoma↗
PubMed2001

Evaluation of formalin-fixed paraffin-embedded tissues from vaccine site-associated sarcomas of cats for polyomavirus DNA and antigen.

OBJECTIVE: To determine whether vaccine site-associated sarcomas (VSS) from cats contain polyomavirus antigen or DNA. SAMPLE POPULATION: 50 formalin-fixed paraffin-embedded tissue blocks of VSS from cats. PROCEDURE: Sections from each tissue block were evaluated for polyomavirus antigen by use of an avidin-biotin-complex immunohistochemical staining method, using rabbit anti-murine polyomavirus polyclonal antiserum as the primary antibody. The DNA was extracted from sections of each tissue block, and a polymerase chain reaction assay was performed, using primers designed to amplify regions of the bovine polyomavirus genome and consensus polyomavirus primers designed to detect unknown polyomaviruses. RESULTS: Polyomavirus antigen and DNA were not detected in any of the VSS. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that polyomaviruses likely do not have any direct involvement in the pathogenesis of VSS in cats.

Animals↗
PubMed2001

Evaluation of formalin-fixed paraffin-embedded tissues from vaccine site-associated sarcomas of cats for papillomavirus DNA and antigen.

OBJECTIVE: To determine whether vaccine site-associated sarcomas (VSS) from cats contain papillomavirus antigen or DNA. SAMPLE POPULATION: 50 formalin-fixed paraffin-embedded tissue blocks of VSS from cats. PROCEDURE: Sections from each tissue block were evaluated for papillomavirus antigen by use of an avidin-biotin-complex immunohistochemical staining method, using rabbit anti-bovine papillomavirus type-1 antibody. The DNA was extracted from sections of each tissue block, and polymerase chain reaction assays were performed, using primers designed to amplify regions of the E5 gene of bovine papillomavirus and consensus primers designed to amplify a region of the L1 gene of animal papillomaviruses. Sections from 20 of the tissue blocks were evaluated by use of nonradioactive in situ hybridization for bovine papillomavirus DNA. RESULTS: Papillomavirus antigen and DNA were not detected in any of the VSS. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that papillomaviruses likely do not have any direct involvement in the pathogenesis of VSS in cats.

Animals↗
PubMed1999

NCL-CD10-270: a new monoclonal antibody recognizing CD10 in paraffin-embedded tissue.

CD10 (CALLA) antigen is expressed in a wide variety of epithelial and nonepithelial tissues, but its most significant application is in the diagnosis and classification of certain types of malignant lymphoma and leukemia. CD10 is expressed in a high percentage of cases of acute lymphoblastic leukemia (ALL), follicular lymphoma, Burkitt's lymphoma, and some hematopoietic tumors. Although the antigen is not lineage specific, CD10 expression is widely used to define subgroups within B-ALL and is a useful tool for detecting the presence of leukemic blasts in the bloodstream. Currently available monoclonal antibodies to CD10 have been found to be effective only in fresh-frozen tissue and for techniques such as flow cytometry. We have used a recombinant protein corresponding to the whole of CD10 to generate a monoclonal antibody that is effective in paraffin-embedded tissue sections. We have used this antibody to assay for the presence of CD10 on a range of normal and pathological tissues. Strong staining was seen in lymphoid germinal centers, renal tubules, glomeruli, syncytiotrophoblast, hepatic parenchymal canaliculi, B-lineage ALL, follicle center cell lymphoma, and a proportion of cases of large-B-cell lymphoma. We believe that this antibody will be of value in the characterization of malignant lymphoma, in particular the differential diagnosis of small-B-cell lymphoma and subtyping of lymphoblastic leukemia, as well as the investigation of the significance of expression of CD10 in other normal and pathological tissues.

Animals↗
PubMed1986

Bone metastases of differentiated and medullary thyroid gland carcinomas. Usefulness and limitations of immunohistology performed on undecalcified plastic-embedded tissue specimens.

Undecalcified methylmethacrylate(MMA)-embedded biopsies and surgical specimens from 20 bone metastases of differentiated or medullary thyroid carcinomas or prostate carcinomas were investigated immunohistologically for the presence of thyroglobulin, cytokeratin, vimentin, and CEA. The immunoreactions on MMA-sections revealed the same staining patterns as those demonstrated using paraffin sections of the primary lesions. Conversely, immunohistological examination of decalcified paraffin-embedded specimens of the same metastases yielded either false-negative results or results that did not allow an exact evaluation. The findings demonstrate the usefulness and limitations of immunohistology when performed on undecalcified plastic-embedded material.

Adult↗
PubMed1986

Immunocytochemical methods for demonstrating canine distemper virus antigen in aldehyde-fixed paraffin-embedded tissue.

The effects of enzymatic digestion, sodium borohydride reduction, acids used in decalcification procedures and techniques for inactivation of endogenous peroxidase were sequentially evaluated for their effect on the immunoreactivity of canine distemper virus in aldehyde-fixed paraffin-embedded tissue. Enzyme digestion improved immunoreactivity while sodium borohydride reduced background staining. Paraformaldehyde-glutaraldehyde-fixed tissues required thioglycolic acid treatment prior to enzyme digestion and sodium borohydride reduction to obtain results comparable to results obtained in formalin-fixed tissues. Detailed protocols for indirect immunofluorescence and the avidin-biotin-peroxidase complex procedure are provided.

Acids↗
PubMed1986

Identification of Brucella abortus in formalin-fixed, paraffin-embedded tissues of cows, goats, and mice with an avidin-biotin-peroxidase complex immunoenzymatic staining technique.

An avidin-biotin-peroxidase complex immunoenzymatic staining technique was evaluated for light microscopic detection of Brucella organisms in formalin-fixed, paraffin-embedded tissues. Tissues from cows, goats, and mice inoculated with B abortus strain 2308 were examined, using rabbit antiserum to Brucella cell surface protein as primary antibody. Stained organisms were identified histologically in tissue sections containing B abortus, as detected by bacteriologic examination of duplicate nonprocessed tissue samples. Bacillus sp, Corynebacterium pyogenes, Escherichia coli, Listeria monocytogenes, Pasteurella haemolytica, P multocida, Salmonella typhimurium, Staphylococcus sp, and Streptococcus sp did not stain with this system, using anti-Brucella cell surface protein primary antibody, thus indicating specificity for Brucella organisms.

Animals↗
PubMed1995

Analysis of K-ras gene mutations in periampullary cancers, gallbladder cancers and cholangiocarcinomas from paraffin-embedded tissue sections.

Point mutations of the K-ras gene were analyzed in 25 periampullary cancers (21 ampulla vater cancers, two common bile duct cancers and two duodenal cancers), two gallbladder cancers and six cholangiocarcinomas. DNA extracted from the paraffin-embedded tissues was amplified with the polymerase chain reaction and subsequently analyzed by direct cycle sequencing at codons 12, 13 and 18 of the K-ras gene. Codon 61 was first screened with single strand conformation polymorphism and then sequenced by direct cycle sequencing. No point mutation was found in any of the 25 periampullary cancers or the two gallbladder cancers. These results are similar to previous reports. Mutation of the K-ras gene seems not to play an important role in tumorigenesis of periampullary cancer. In two of six (33%) cholangiocarcinoma patients, point mutations were found. Both mutations were transitions, GGT to GAT at codon 12. The incidence of mutation was greater than that in Thailand (about 8%) but less than that in Japan (about 60%). Mutation of the K-ras gene may play varied roles in the tumorigenesis of cholangiocarcinoma, depending on geographic area.

Adult↗