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

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

OBJECTIVE: To explore the method for DNA typing in formalin-fixed tissue by detecting SNP markers on X chromosome. METHODS: Genomic DNA was prepared from formalin-fixed tissue. In the event that typing using Sinofiler and MiniFiler kits had failed, 51 SNPs were amplified with multiplex PCR and typed with matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). RESULTS Full profiles of X-SNP loci were obtained from the formalin-fixed tissue, while the STR and miniSTR genotyping failed. CONCLUSION SNP genotyping technique can be used to obtain more information for formalin fixed tissues.
Chromosomes, Human, X ; DNA/isolation & purification* ; DNA Primers ; Female ; Forensic Genetics ; Formaldehyde ; Genotype ; Humans ; Intestines ; Multiplex Polymerase Chain Reaction ; Paraffin Embedding ; Polymorphism, Single Nucleotide ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Tissue Fixation/methods*

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

OBJECTIVE: To assess the influential factors of STR genotyping in 10% unbuffered formalin fixed paraffin embedded samples. METHODS: Eight kinds of autopsy samples including heart, brain, liver, spleen, kidney, lung, stomach and intestine tissue from 2 corpse were fixed with 10% unbuffered formalin and embedded with paraffin according to the routine procedure from which the DNA were extracted with three different methods (QIAGEN, IQ and Chelex). STR profile were analyzed with AmpFlSTR Identifiler Kit and capillary electrophoresis on genetic analyzer 3100-Avant. STR profiles of 56 archival paraffin embedded samples from 15 cases were also analyzed with methods as mentioned. These archival samples, including heart, liver, lung and intestine tissue, had been preserved for 1 to 5 years in ambient temperature. Effectiveness of STR genotyping was assessed with the recalling ration of the 15 STR loci composing of the Identifiler Kit. RESULTS: Significant difference of the recalling ration was statistically revealed among the different types of paraffin embedded sample with same preserving period. Moreover, the STR recalling ration was continuously lowering with the prolongation of preserving period in all of the samples. The linear relationship between the STR recalling ratio and the preserving period was showed in lung and heart sample. The STR recalling ration in lung sample was higher than that in the other types of paraffin embedded sample. CONCLUSION Preserving period, tissue type, extracting method of DNA and the PCR template concentration were the most important influential factors for successfully STR genotyping paraffin embedded samples, which fixed with unbuffered formalin for the same time.
DNA/isolation & purification* ; DNA Fingerprinting/methods* ; Forensic Genetics/methods* ; Formaldehyde/chemistry* ; Humans ; Liver ; Lung ; Myocardium ; Paraffin Embedding ; Polymerase Chain Reaction ; Specimen Handling/methods* ; Tandem Repeat Sequences ; Time Factors ; Tissue Fixation

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

Adolescent ; Child ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Leg ; Male ; Paraffin Embedding ; Sarcoma, Synovial ; diagnosis ; genetics ; pathology ; Soft Tissue Neoplasms ; diagnosis ; genetics ; pathology ; Translocation, Genetic

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

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