Histocytological Preparation Techniques ; methods ; Microtomy ; methods ; Tissue Embedding

Paraffin Embedding ; methods ; Tissue Array Analysis ; instrumentation ; methods

OBJECTIVE: To optimize the method for embedding multiple undecalcified mouse tibias in plastic blocks, improve the efficiency and stability of plastic embedding and reduce the detachment rate of plastic slides. METHODS: Thirty undecalcified tibias from 15 B6 mice were used for plastic embedding after calcein labeling, fixation, dehydration and infiltration. The tibias were embedded in cylindrical plastic blocks with a diameter of 4 mm. For each bone, the 1/4 proximal tibia was cut off, and the remaining 3/4 was used for re-embedding. Five bones were embedded in a single block with each bone standing closely on the surface of a flat plate. The samples were randomized into control and experimental groups in all the processes of embedding, sectioning and staining. In the 3 groups with modified embedment, flowing CO was added into the embedding solution, embedding solution was applied to the section surface, and the slides were heated at 95 ℃ for 15 min. The polymerization time, slide detachment rate, bone formation and osteoblast parameters were analyzed. RESULTS: We prepared 6 plastic blocks, each containing 5 tibias, whose cross sections were on the same plane. The blocks were completely polymerized and suitable for sectioning. Flowing CO into the embedding solution reduced the polymerization time and increased the rate of complete polymerization. Application of the embedding solution on the section surface significantly reduced the detachment rate of the sections ( < 0.05) without affecting bone formation analysis ( > 0.05). Heating the slides significantly lowered the detachment rate of the sections ( < 0.05) without affecting osteoblast analysis ( > 0.05). CONCLUSIONS The optimized method allows effective embedding of multiple undecalcified mice tibias in the same block and can be an ideal method for histological analysis of undecalcified bones.
Animals ; Mice ; Plastics ; Staining and Labeling ; Tibia ; Tissue Embedding ; methods

Formaldehyde ; Humans ; Microdissection ; Paraffin Embedding ; Proteomics ; methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Tissue Fixation

Antigens ; analysis ; genetics ; Fixatives ; Formaldehyde ; Hot Temperature ; Humans ; Immunohistochemistry ; methods ; In Situ Hybridization ; methods ; Paraffin Embedding ; Proteins ; analysis ; genetics ; Staining and Labeling ; methods ; Tissue Fixation ; methods

Antigens ; analysis ; immunology ; Antigens, CD ; analysis ; immunology ; Formaldehyde ; Hot Temperature ; Humans ; Immunohistochemistry ; methods ; Neoplasms ; metabolism ; pathology ; Paraffin Embedding ; methods ; Staining and Labeling ; Tissue Fixation ; methods

Adult ; Aged ; Aged, 80 and over ; Female ; Gastritis, Atrophic ; diagnosis ; pathology ; Gastroscopy ; methods ; standards ; Humans ; Male ; Middle Aged ; Specimen Handling ; methods ; Tissue Embedding ; methods

Owing to the DNA degradation induced by formalin and the obstruction of paraffin to DNA extraction, it is difficult to recover high-quality DNA from Formalin fixed and paraffin embedded tissue (FFPET). In recent years, a lot of researches indicate that the DNA extraction from FFPET can be developed by improving the pretreatment, optimizing the digestion condition of proteinase, simplizing the procedures of the DNA extraction, purifying the extracted DNA and so on, which may pave a way for popularizing FFPET in DNA analysis.
DNA/isolation & purification* ; Forensic Medicine/methods* ; Formaldehyde/chemistry* ; Humans ; Hydrogen-Ion Concentration ; Paraffin Embedding/methods* ; Polymerase Chain Reaction ; Specimen Handling/methods* ; Time Factors ; Tissue Fixation

The CDKN2 (MTS1/p16INK4A) gene, encoding cyclin dependent kinase inhibitor, was found to be homozygously deleted at a high frequency in cell lines from many different types of cancer and some primary cancers. To determine the frequency of CDKN2 mutations in most common human cancers in Korea, PCR and PCR-SSCP analyses for the exon 2 of CDKN2 were performed on each set of 20 formalin-fixed and paraffin-embedded tumor tissues of stomach adenocarcinomas, lung cancers, cervix cancers and hepatocellular carcinomas. No mutations in exon 2 of CDKN2 were found in 20 stomach adenocarcinomas. In contrast to rare mutations in stomach adenocarcinomas, a high frequency of CDKN2 mutations was identified in other 3 cancers, 11 of 20 (55%) lung cancers (7 of 10 NSCLCs and 4 of 10 SCLCs), 14 of 20 (70%) cervix cancers and 11 of 20 (55%) hepatocellular carcinomas. These results suggest that mutations of the CDKN2 gene might be an important genetic change in NSCLCs, cervix cancers and hepatocellular carcinomas.
Adenocarcinoma/genetics ; Carcinoma, Hepatocellular/genetics ; Cervix Neoplasms/genetics* ; Female ; Formaldehyde ; Human ; Liver Neoplasms/genetics* ; Lung Neoplasms/genetics* ; Mutation ; Paraffin Embedding ; Protein p16/genetics* ; Sequence Deletion ; Stomach Neoplasms/genetics* ; Tissue Embedding/methods

The aim of this research was to explore the most optimal method of DNA extraction from formalin-fixed, paraffin-embedded (FFPE) tissues, and to improve the amplification of long fragments with the method. Three methods, one step method, phenol-chloroform extraction method, and genomic DNA purification kit method, were employed to extract DNA from twenty normal thyroid tissues which were fixed with formalin and embedded with paraffin. The highest proportionality of OD260/OD280 in the examples was obtained by phenol-chloroform extraction method, 1.703 +/- 0.086, compared to the results of the other two methods. As for the long DNA segments amplification, the achievement ratio of one step method, phenol-chloroform extraction method and genomic DNA purification kit method were 0%, 5% and 10%, respectively, by traditional PCR method, but 0%, 95% and 85% respectively by the nest PCR. We have found that the best process of extracting DNA from FFPE is digesting by proteinase K and purifying by phenol-chloroform, and it is effective to amplify long DNA segments from FFPE by nest PCR.
Base Sequence ; DNA ; isolation & purification ; Formaldehyde ; Humans ; Molecular Sequence Data ; Paraffin Embedding ; Pathology, Molecular ; methods ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Thyroid Gland ; pathology ; Tissue Fixation