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 307 records · Page 17Linked to original sources
PubMed1983

Detection of viral genomes in cultured cells and paraffin-embedded tissue sections using biotin-labeled hybridization probes.

A method of in situ cytohybridization is described for the detection of specific viral genomes in infected cell cultures or paraffin-embedded tissue sections without the use of radioisotopes. Biotin-labeled analogs of TTP are incorporated into viral DNA in vitro by nick translation and the resultant DNA probes hybridized to cytologic samples. Cells containing viral genetic material are then revealed by standard immunofluorescence, immunoperoxidase, or affinity cytochemical techniques that are based on the specific interaction between biotin and antibiotin IgG or avidin. Hybridization probes containing nucleotides that have an 11- or 16-atom spacer arm between the biotin molecule and the pyrimidine ring interact with these detector proteins more efficiently than probes containing biotin-nucleotides with a 4-atom spacer arm. The total procedure can be performed fairly rapidly (24 hr or less) and numerous samples can be processed simultaneously. Although the detection methods employed to date are not as sensitive as autoradiographic procedures with high specific activity probes, more sensitive protein detector complexes are currently being constructed. The speed, specificity, and resolving power of this technique should be of general utility in screening for the presence of infectious agents in cell or tissue samples. Here we report the visualization of parvovirus, polyomavirus, herpes simplex virus, adenovirus, and retrovirus genetic material in infected cell cultures and herpes simplex and adenovirus DNA in paraffin-embedded autopsy tissues.

Adenoviruses, Human↗
PubMed1995

Localization of Epstein-Barr Virus-Encoded Small RNA-1 by in situ Reverse Transcription: Demonstration of cDNA Generation in Formalin-Fixed Paraffin-Embedded Tissue Sections.

Reverse transcription (RT) followed by polymerase chain reaction (RT-PCR) has been commonly used to detect viral and cellular transcripts in whole cell extracts. Application of this technique to tissue sections requires the in situ generation of cDNA. In this study, we selected an abundant transcript, Epstein-Barr virus (EBV)-encoded small RNA (EBER-1), as a model template to demonstrate cDNA generation in tissue sections. Using both digoxigenin-dUTP and primers which are complementary to EBER-1, we demonstrated specific EBER-1 cDNA generation both in vitro, and in tissue sections taken from formalin-fixed paraffin-embedded cell blocks of an EBV-infected cell line, B95-8. Furthermore, we utilized in situ RT in sections of EBV-associated nasopharyngeal carcinomas, and identified EBER-1 cDNA specifically in neoplastic cells, but not in the surrounding nonneoplastic stroma. EBER-1 cDNA was localized to the nucleus of these cells, with relative sparing of the nucleolus and the cytoplasm. No specific signal was evident if the reverse transcriptase was omitted, if 'sense' primers were used, or if RT was preceded by RNase digestion. The specificity of EBER-1 cDNA was further confirmed by in situ hybridization using the sense riboprobe, which has the same polarity as the EBER-1 transcript. Our results provide a successful example of using nonradioactive nucleotide analogue for cDNA generation in formalin-fixed, paraffin-embedded tissue sections. This approach would provide a visible assay to monitor RT in tissue sections, and allow further optimization of conditions for cDNA generation in tissue sections. Therefore, it potentially can be helpful for the future development of RT-PCR in tissue sections. Copyright 1995 S. Karger AG, Basel

Journal Article↗
PubMed1978

A reliable method for electron microscopic examination of specific areas from paraffin-embedded tissue mounted on glass slides.

A reliable method for electron microscopic examination of specific areas from paraffin-embedded tissue mounted on glass slides. Am J Clin Pathol 70: 697--699, 1978. A method to embed and cut tissue mounted on ordinary glass histologic preparations for electron microscopic observations is described. The cover slip is removed by immersing the slide in xylene for a few minutes to several hours. After the cover slip is removed, the tissue is fixed in osmium tetroxide, dehydrated, and embedded in epoxy resins by inverting a BEEM capsule filled with the resin on a selected area of the slide. After polymerization, thin sections are obtained and observed with the electron microscope with good results.

Histological Techniques↗
PubMed1980

A rapid method for embedding tissues for electron microscopy using 1,4-dioxane and Polybed 812.

A rapid method for embedding tissues for electron microscopy is described. This method, which can be completed within 5 hr, uses 1,4-dioxane as the final dehydrating agent and Polybed 812 as the embedding medium. Satisfactory preservation of cellular structures is consistently achieved with a variety of normal and diseased tissues. This method may be of particular value to diagnostic electron microscopy laboratories where time and simplicity are critical.

Animals↗
PubMed1984

DNA-Feulgen-cytophotometric analysis of single cells isolated from paraffin embedded tissue.

This study describes a method for the isolation of cells from paraffin embedded tissues for DNA-Feulgen-cytophotometric measurements. The relevance of the method used is demonstrated by the analysis of the DNA-distribution pattern of cells isolated from an alveolar sarcoma of the soft tissues of a 10-year old girl. The comparison between freshly prepared imprint preparations and specimens after pepsin--extraction clearly shows, that the preparation mode used did not influence the relative DNA-content. The mean values of DNA/nuclei did not show a decrease even after prolonged pepsin treatment (90 minutes). Therefore this method permits access to stored histopathological material for retrospective DNA-Feulgen-Cytophotometric investigations.

Child↗
PubMed1978

A method for specific diagnosis of Rocky Mountain spotted fever on fixed, paraffin-embedded tissue by immunofluorescence.

For specific demonstration of Rickettsia rickettsii in fixed, paraffin-embedded tissues, the technique of trypsin digestion of deparaffinized, rehydrated sections was investigated. It was determined that 3.5 hr of digestion was optimal for achieving discrete, bright, green immunofluorescence of rickettsiae. At autopsy kidneys from seven of 10 cases of probably Rocky Mountain spotted fever contained structures that were specifically stained and that had the size and shape of rickettsiae. These structures were strictly limited to the endothelium and vascular walls of renal capillaries, veins, and arteries. Results from controls indicated that specificity of the immunofluorescence. R. rickettsii may be demonstrated by this method with greater sensitivity and specificity than by current histological methods. The technique allows retrospective analysis of certain organs for pathogenesis of involvement in Rocky Moutain spotted fever and offers a specific diagnostic test.

Animals↗
PubMed1982

A grinding method for producing sections of large areas of plastic embedded tissues.

We have developed a method utilizing relatively thick ground sections of plastic embedded tissue which affords the resolution obtained with 0.5 micrometers cut sections. The sections, which are permanently affixed to plastic microscope slides, are much larger in area than ultramicrotome sections. Additional advantages are: sections can be destained and restained and selected areas can be examined with various forms of electron microscopy. Autoradiographic studies are also possible. Although the method has a broader application, it is particularly useful in examining the interface between hard and soft tissues.

Animals↗
PubMed1990

DNA ploidy analysis of endometrial adenocarcinoma using a flow cytometry from paraffin-embedded tissues.

In this study, DNA ploidy using a flow cytometry from paraffin-embedded tissues was evaluated to establish the DNA heterogeneity and risk factor of endometrial adenocarcinoma. Seventy-six samples from 31 patients with endometrial adenocarcinoma and 6 controls with normal endometrium were measured for DNA content. DNA ploidy of endometrial adenocarcinoma was also analyzed in three different sites of a tumor, viz. (1) the primary endometrial lesion, (2) the site of myometrial invasion and, (3) metastatic lesions. In over 22% of 27 patients with endometrial adenocarcinoma, intratumor variations in DNA ploidy were demonstrated of a different sites of a tumor. It was found that all 6 controls with normal endometrium were diploid, while 28.6% of 70 samples of endometrial adenocarcinoma had aneuploid tumors. Poorly differentiated tumors had a significantly higher incidence of aneuploidy than well or moderately differentiated tumors. DNA ploidy was related to the surgical stage and histological grade, although it was not indicated that DNA ploidy has reliable prognostic value for endometrial adenocarcinoma. It was suggested that the myometrial invasion of endometrial adenocarcinoma is not an important factor in DNA content heterogeneity, because the incidence of aneuploidy in three different sites of a tumor was as follows: myometrial invasion less than primary endometrial lesion less than metastatic lesions.

Adenocarcinoma↗
PubMed1998

How thick are the paraffin-embedded tissue sections routinely prepared in laboratory? A morphometric study using a confocal laser scanning microscope.

In order to evaluate the differences between cut and measured thicknesses of formalin-fixed, paraffin-embedded tissue sections, formalin-fixed, paraffin-embedded blocks of liver tissue were prepared from four male Wistar rats. The sections were cut at preselected thicknesses of 3, 6, 9, 12, 15 and 18 microns, and observed under confocal laser scanning microscopy. The maximum and minimum section thicknesses were measured using a vertically cut section image constructed by the software incorporated in the microscope. The surface of the section was not smooth but showed fine undulations in addition to a slight inclination. The mean and range of the section thicknesses measured were 4.3 +/- 1.3, 7.0 +/- 1.7, 9.8 +/- 2.1, 12.0 +/- 2.6, 14.1 +/- 3.0, and 17.7 +/- 3.8 microns for the preselected cut thicknesses of 3, 6, 9, 12, 15 and 18 microns. It is stressed that the evaluated parameters of the section (volume or surface densities) include standard errors of up to 10% caused by the differences between the cut and real thicknesses.

Animals↗
PubMed2004

Chemical characterization of a lambda I amyloid protein isolated from formalin-fixed and paraffin-embedded tissue sections.

Amyloid protein was isolated from formalin-fixed paraffin-embedded heart tissue sections from a patient with primary (AL) amyloidosis by extraction with 6 M guanidine HCl. SDS-PAGE analysis of extracted material showed a major band at 16 kDa and a minor band at 18 kDa. Edman degradation analysis before and after pyroglutamate aminopeptidase treatment showed that the amyloid protein contained N-terminal pyroglutamic acid and was derived from an immunoglobulin lambda light chain. Analysis of tryptic peptides from the extract identified the amyloid protein as a lambda I. Of particular interest is that almost the entire amyloid protein amino acid sequence could be obtained from the cardiac sections. These results demonstrate that formalin-fixed paraffin-embedded tissue sections can be used for extensive biochemical characterization of amyloid proteins and will become a valuable source for isolation and extensive biochemical characterization of amyloid proteins as they are now a valuable source for isolation of DNA for genetic analysis.

Amino Acid Sequence↗
PubMed1992

Comparison of eight modifications of Hedley's method for flow cytometric DNA ploidy analysis of paraffin-embedded tissue.

Flow cytometric DNA analysis of nuclear suspensions from formalin-fixed, paraffin-embedded tissue often fails to detect aneuploid cell populations present in corresponding fresh tissue. Nuclear suspensions were prepared by 8 different modifications and standard Hedley's method using 50-microns sections of tissue blocks from 8 breast and 8 colonic carcinomas, all previously known to be DNA aneuploid by analysis of fresh tumor. Pepsin solutions of three different enzymatic activities were used to release nuclei using three different tissue digest formats. DNA aneuploidy was demonstrated overall in 7 of 72 different colon tumor experiments and 25 of 72 breast cancer experiments. Modifications yielded aneuploid populations not detected by the standard Hedley method; DNA aneuploidy of 4 breast and 2 colon cancers was detected by modifications compared to 2 breast and 1 colon cancer demonstrated by the standard. No single method consistently demonstrated DNA aneuploidy. High histogram baselines, presumably from debris, contributed to the marked loss of sensitivity in detecting most DNA aneuploid populations. Detection of DNA aneuploidy was most closely associated with specific cases, regardless of the method of nuclear suspension preparation. Recovery of DNA aneuploid nuclei seems to depend primarily on tissue processing or innate characteristics of the tumor cells, not on the method used to prepare the nuclear suspension.

Breast Neoplasms↗
PubMed1995

Extramedullary myeloid cell tumors. An immunohistochemical study of 29 cases using routinely fixed and processed paraffin-embedded tissue sections.

OBJECTIVE: Extramedullary myeloid cell tumors (EMCTs) may be unsuspected clinically and difficult to recognize histologically. Fresh or frozen tissue is often not available for analysis. We studied 29 cases of EMCT using routinely fixed and processed paraffin-embedded tissue, an immunohistochemical method, and a panel of antibodies. PATIENTS: We studied 29 patients with EMCTs: 22 males and 7 females, with a median age of 48 years (range, 5 to 80 years). Histologically, 9 tumors were well differentiated, 16 were poorly differentiated, and 4 were blastic. RESULTS: The Leder stain (napthol-ASD-chloroacetate esterase) was positive in 21 (77.7%) of 27 tumors. Immunohistochemically, the following antibodies reacted with the greatest number of cases: Leu-22 or MT1 (CD43) in 28 (96.6%) of 29, antilysozyme in 27 (96.4%) of 28, and antimyeloperoxidase (MP07) in 21 (91.3%) of 23 cases. Other myeloid lineage-associated antibodies were positive in a subset of cases: antineutrophil elastase (NP57) in 10 (62.5%) of 16, Leu-M1 (CD15) in 7 (46.6%) of 15, and Mac-387 in 6 (40.0%) of 15 cases. The well-differentiated EMCTs reacted with most myeloid-associated antibodies; poorly differentiated and blastic tumors were more often negative. The pan-leukocyte antibody LCA (CD45RB) reacted with 15 (60%) of 25 neoplasms. Three (16.6%) of 18 tumors contained numerous p53-positive cells, ranging from 10% to 50% of the tumor cell population. In 10 cases, exons 5 through 8 of the p53 gene were analyzed using the polymerase chain reaction and single-stranded conformational polymorphism analysis. Gel shifts consistent with mutations were identified in exon 8 of one tumor (10%) that exhibited abundant p53 immunostaining. CONCLUSIONS: Immunohistochemical studies using fixed, paraffin-embedded sections are very useful in the diagnosis of EMCTs. The most sensitive antibodies are anti-CD43, antilysozyme, and antimyeloperoxidase. Immunohistochemical methods are more sensitive than the Leder stain. We found p53 staining in a small subset of cases, in which we were able to confirm evidence of p53 gene mutation using the polymerase chain reaction and single-stranded conformational polymorphism analysis in one case; p53 gene mutations appear to be uncommon in EMCTs.

Adult↗
PubMed2002

[Investigation of practical application of fluorescence in situ hybridization (FISH) analysis using microwave irradiation in formalin-fixed, paraffin-embedded tissue sections].

We investigated fluorescence in situ hybridization (FISH) analysis using microwave irradiation in formalin-fixed, paraffin-embedded tissue sections of breast fibroadenoma. Higher percentage of cells with 2 signal copies of chromosome 3 centromere could be obtained in the condition of 5 microns thick sections, when we counted cells of more than 4 microns of nuclei in thickness. This method showed about the same results as FISH using cells separated from the same tissues. Percentage of cells with 2 signal copies of chromosome 17 centromere in 14 cases was 80.6 +/- 4.0% (Mean +/- S.D.). This method is expected in the application of the prognosis estimation of the breast cancer.

Breast Neoplasms↗
PubMed1990

Enzyme histochemistry on freeze-substituted glycol methacrylate-embedded tissue.

We developed a method for histochemical demonstration of a wide range of enzymes in freeze-substituted glycol methacrylate-embedded tissue. Tissue specimens were freeze-substituted in acetone and then embedded at low temperature in glycol methacrylate resin. All enzymes studied (oxidoreductases, hydrolases) were readily demonstrated. The enzymes displayed high activity and were accurately localized without diffusion when tissue sections were incubated in aqueous media, addition of colloid stabilizers to the incubating media not being required. Freeze-substitution combined with low-temperature glycol methacrylate embedding permits the demonstration of a wide range of enzymes with accurate enzyme localization, maintenance of enzyme activity, and excellent tissue morphology.

Acrylates↗
PubMed1984

Immunoperoxidase staining for Ia-like antigens in paraffin-embedded tissues from human melanoma and lung carcinoma.

The human Ia-like antigens that are predominantly expressed by cells associated with immunologic function has been considered as a diagnostic marker of malignant transformation of some nonlymphoid tissues. Immunoperoxidase staining of formalin-fixed and paraffin-embedded tissue sections with a monoclonal antibody to Ia-like antigens was chosen for assessment of the value of this marker for diagnosis in surgical pathology. Monoclonal antibody LK8D3 developed against a human melanoma cell line bearing Ia-like antigens was found to react in serologic and immunochemical studies with an antigenic determinant of Ia-like antigens that was relatively stable to formalin fixation and paraffin embedding. Avidin-biotin complex peroxidase staining of formalin-paraffin sections with LK8D3 showed focal expression of Ia-like antigens in 3 of 12 melanomas, whereas all 8 cases of intradermal nevi were negative. Immunoperoxidase staining of formalin-paraffin sections of lung carcinomas with antibody LK8D3 was related to the histologic subtype of tumors. Thus, squamous cell carcinomas showed only very focal staining for Ia-like antigens in 5/9 cases, while widespread and intense Ia-like immunoreactivity was seen in 3/5 cases of lung adenocarcinomas, including two bronchioalveolar carcinomas. The presence of Ia-like antigens in lung adenocarcinoma may not be entirely associated with malignant transformation, because normal alveolar lining cells were stained with the antibody.

Adenocarcinoma↗
PubMed1991

Gold toning improves the visualization of nucleolar organizer regions in paraffin embedded tissues.

A modification of the silver colloid technique for staining nucleoar organizer regions in paraffin embedded tissues is described. This modification involves the application of a gold toning step with subsequent gold reduction, if necessary, following incubation of sections in the standard silver colloid solution. Silver stained nucleolar organizer regions (AgNORs) in toned sections are more sharply delineated when compared to untoned controls. In high grade tumors the addition of the toning step results in significantly higher AgNOR counts due to the ability to discriminate more easily individual AgNORs in argyrophilic aggregates within the nucleus. It is recommended, because of enhanced visualization, that this modification of the silver colloid technique be used in studies involving quantification of AgNORs in tissue sections.

Gold↗
PubMed1992

DNA content in fresh versus paraffin-embedded tissue. Flow cytometric analysis of 100 tumors.

DNA ploidy analysis was determined on 100 consecutive tumors from a wide variety of sites using both fresh and paraffin-embedded tissue on the same specimen. The correlation coefficient (r) value between the methods was 0.85. Aneuploidy was detected by both methods in 51/100 (51%) of the cases. Fresh tissue analysis yielded 10 additional cases (overall 61% aneuploidy) not detected on corresponding paraffin-embedded sections, whereas paraffin-embedded analysis detected 4 additional cases (overall 55% aneuploidy) not revealed by fresh tissue analysis. Fresh tissue analysis produced lower coefficients of variation and resulted in a cleaner preparation with less cellular debris. Fresh tissue analysis was also superior to paraffin for the detection of hypodiploid, near-diploid and multiple peaks. Analysis of paraffin-embedded material allows examination of archival tissue and provides a more rapid means of long-term follow-up and statistical correlations for prognostic studies. Although the overall correlation of both methodologies for DNA analysis showed a minimal variation in results, in our experience fresh tissue analysis has an advantage and is preferable, when available, for ploidy analysis.

Cell Separation↗