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PubMedDate not supplied

Southern blot analysis of DNA extracted from paraffin wax-embedded tissue.

Genetic analysis of multi-generation families using RFLP is often incomplete because fresh tissue is not available for the isolation of DNA. However, in many instances archival material exists, usually formalin-fixed in paraffin blocks. Procedures have been described for extracting DNA from such specimens. Many parameters affect the quality of the DNA eventually extracted and this in turn determines whether the DNA can be successfully digested with restriction enzymes and probed (Dubeau et al., 1986; Warford et al., 1988). In essence, the procedures followed at the time of fixation largely determine whether archival material will yield useful results. We have re-examined the effect of varying the fixation time but, more importantly, we have assessed the validity of RFLP results obtained from fixed tissue.

Blotting, Southern↗
PubMed1991

Enhancement of immunoreactivity among lymphoid malignant neoplasms in paraffin-embedded tissues by refixation in zinc sulfate-formalin.

It has been previously demonstrated that improved cytomorphologic detail can be obtained from tissue blocks initially fixed in 4% formaldehyde solution by means of a "run-back" procedure to an aqueous phase of solution, allowing refixation with a protein-precipitating agent, such as zinc sulfate-4% formaldehyde solution. Because anecdotal experience with immunostaining such reprocessed tissues had suggested improved results, we performed a prospective trial involving 13 cases to compare the intensity of immunoreactivity before and after such reprocessing. In 10 of 13 cases studied with a variety of antibody reagents, we noted a definite improvement of staining, preferentially among those with the weakest original immunoreactivity.

Antibodies, Neoplasm↗
PubMed1990

Detection of canine distemper viral antigen in formalin-fixed and paraffin-embedded tissue of a fitch (Mustela putorius), using an immunoperoxidase technique.

The present study describes how a naturally infected fitch (Mustela putorius) caused an outbreak of canine distemper in a colony of insufficiently vaccinated dogs. The detection of canine distemper virus (CDV) on paraffin sections of different organs of the fitch and one of the dead dogs was achieved using a monoclonal antibody against the nucleocapsid protein (NP) of CDV and the avidin-biotin-peroxidase complex (ABC) technique.

Animals↗
PubMed1986

Oxidized p-phenylenediamine: observations on the staining reaction in epoxy embedded tissues.

p-Phenylenediamine (PPD) is easily oxidized to brown compounds which stain acidic substrates. On account of the spontaneous oxidation process, the colour of PPD increases and becomes ochre-brown in a few days, showing an absorption peak at lambda = 510 nm with shoulder at about 440 to 460 nm. Studies on the application of oxidized PPD as a stain for semi-thin sections revealed that some tissue components could be clearly visualized. After glutaraldehyde fixation, semi-thin and thin sections of animal tissues were treated with 0.5% aqueous PPD solutions which were aged for variable times at room temperature. Microvilli, goblet cell mucin, mast cell granules, cartilage matrix, collagen, elastin, keratohyalin granules, acrosomes, cytoplasmic granules of Drosophila hydei salivary glands and chromatin showed positive staining reactions after treatment of semi-thin sections with oxidized PPD (7-10 days aged) for 20-30 minutes. Microspectrophotometric studies revealed an absorption peak at lambda = 520-530 nm and a shoulder at lambda = 440-460 nm in goblet cell mucin stained by oxidized PPD. In the presence of anionic macromolecules, the main peak of oxidized PPD solutions showed a strong hyperchromism. Thin sections stained by oxidized PPD did not appear contrasted, but the treatment with 0.125% gold chloride (AuCl3) induced massive gold deposits in structures stained by oxidized PPD. Hyperchromic shifts were also produced in oxidized PPD solutions after the addition of small amounts of AuCl3. This procedure can be used as a simple and rapid staining method for epoxy sections, giving selective contrast for some tissue components.

Animals↗