Articles de revues : « Fixed and Paraffin Embedded Tissues »

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Giusti, Laura, et Antonio Lucacchini. « Proteomic studies of formalin-fixed paraffin-embedded tissues ». Expert Review of Proteomics 10, n^o 2 (avril 2013) : 165–77. http://dx.doi.org/10.1586/epr.13.3.

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Díaz-Cano, Salvador J., et Stephen P. Brady. « DNA Extraction from Formalin-fixed, Paraffin-embedded Tissues ». Diagnostic Molecular Pathology 6, n^o 6 (décembre 1997) : 342–46. http://dx.doi.org/10.1097/00019606-199712000-00006.

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Liapis, Helen, et Karen Hutton. « Detection of Integrins in Formalin-fixed, Paraffin-embedded Tissues ». Journal of Histochemistry & ; Cytochemistry 45, n^o 5 (mai 1997) : 737–41. http://dx.doi.org/10.1177/002215549704500512.

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Integrins are heterodimeric transmembrane receptors, which are expressed in many cells. In vitro experiments have demonstrated that integrins may be important in tumor progression and organ development. The functions of integrins were previously studied in cell cultures and their tissue expression was detected by immunofluorescence or immunoperoxidase in frozen sections. The purpose of this study was to determine the optimal conditions for detection of integrins in formalin-fixed, paraffin-embedded tissues. We utilized microwave heating and enzyme digestion in routinely processed, surgically removed tissues. Our results demonstrate that integrins can be reliably detected in archival material. This approach will facilitate further investigation of the role played by integrins in human malignancies and in developmental processes.

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Kerns, B. J., P. A. Jordan, M. B. Moore, P. A. Humphrey, A. Berchuck, M. F. Kohler, R. C. Bast, J. D. Iglehart et J. R. Marks. « p53 overexpression in formalin-fixed, paraffin-embedded tissue detected by immunohistochemistry. » Journal of Histochemistry & ; Cytochemistry 40, n^o 7 (juillet 1992) : 1047–51. http://dx.doi.org/10.1177/40.7.1607637.

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Mutation and overexpression of the p53 gene have been noted in a wide range of human cancers and are thought to play a role in malignant transformation. Previously, immunohistochemical detection of p53 has been possible only in fresh-frozen tissues. We examined p53 expression in paraffin-embedded tissues from 50 epithelial ovarian cancers and 25 primary breast cancers with a modified immunohistochemical (IHC) technique developed in this laboratory, using monoclonal antibody (MAb) PAb1801. The 75 cases were selected from a group of patients in whom the expression levels had already been assessed in a fresh-tissue IHC assay. An identical staining reactivity was observed in both formalin-fixed, paraffin-embedded tissue and fresh-frozen tissue in 48 of 50 (96%) epithelial ovarian cancers and in 23 of 25 (92%) primary breast cancers. Immunodetection of p53 in paraffin-embedded tissue blocks will be a useful alternative to the standard fresh-tissue assay and can accurately reflect the level of p53 expression in human tumors.

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Beckstead, J. H. « A simple technique for preservation of fixation-sensitive antigens in paraffin-embedded tissues. » Journal of Histochemistry & ; Cytochemistry 42, n^o 8 (août 1994) : 1127–34. http://dx.doi.org/10.1177/42.8.8027531.

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Immunohistochemistry is a powerful tool for tissue diagnosis and research. Although the frozen section has remained the gold standard for this important approach to evaluation of antigens in tissues, there is widespread acknowledgment of many limitations. Routine paraffin-embedded sections ware widely used for morphological examination of tissues but are not optimal for antigen preservation. In this study, paraffin-embedded tissues fixed with a simple buffer containing zinc as the primary fixative were compared with tissues fixed with routine formalin, zinc-formalin, paraformaldehyde, ethanol, a variety of commercial (non-formalin-containing) fixatives that have been recommended for reduced toxicity and improved antigen survival, and frozen sections. Human lymphoid tissues and a group of antibodies to antigens (CD1, CD4, CD7, CD8, CD19) usually preserved only in frozen tissue were used as a model system. Fixation in a simple solution of zinc acetate and zinc chloride in a Tris-Ca acetate buffer resulted in antigen preservation comparable to that in frozen sections with antibodies to these cell surface markers. Morphological preservation was comparable to formalin-fixed sections. The work presents a new method that represents the closest approach yet to a technique that combines optimal antigenic survival with the convenience and morphological preservation of traditional formalin-fixed tissue embedded in paraffin.

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Whiteland, J. L., S. M. Nicholls, C. Shimeld, D. L. Easty, N. A. Williams et T. J. Hill. « Immunohistochemical detection of T-cell subsets and other leukocytes in paraffin-embedded rat and mouse tissues with monoclonal antibodies. » Journal of Histochemistry & ; Cytochemistry 43, n^o 3 (mars 1995) : 313–20. http://dx.doi.org/10.1177/43.3.7868861.

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We describe a method for immunohistochemical localization of T-cells, CD4+ T-cells, CD8+ T-cells, B-cells, activated lymphocytes, major histocompatibility complex (MHC) class II antigens, macrophages, dendritic cells, and granulocytes in rat and mouse tissue fixed in periodate-lysine-paraformaldehyde (PLP) and embedded in paraffin. Rat and mouse spleen and eyes were fixed in PLP for 18-24 hr, rapidly dehydrated, infiltrated under vacuum with paraffin at 54 degrees C, sectioned, and stained with appropriate monoclonal antibodies (MAbs). Sections of PLP-fixed, paraffin-embedded spleen were compared with acetone-fixed frozen spleen sections with respect to morphology and staining quality. Nine of 10 MAbs to rat antigens and eight of nine MAbs to mouse antigens stained paraffin sections equally or more intensely than frozen sections. The two MAbs that showed weaker staining still gave good staining on paraffin sections. Paraffin-embedded rat and mouse eyes were easier to section serially than frozen eyes, showed superior morphology, and individually stained cells were readily identified. Therefore, a combination of PLP fixation and low-temperature paraffin embedding permits detection of the major types of immune cell in rat and mouse tissues while maintaining good morphology, particularly in diseased, damaged, or delicate tissues.

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Legrand, Beatrice, Philippe de Mazancourt, Michel Durigon, Véronique Khalifat et Karine Crainic. « DNA genotyping of unbuffered formalin fixed paraffin embedded tissues ». Forensic Science International 125, n^o 2-3 (février 2002) : 205–11. http://dx.doi.org/10.1016/s0379-0738(01)00641-7.

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Blonder, Josip, et Timothy D. Veenstra. « Clinical Proteomic Applications of Formalin-Fixed Paraffin-Embedded Tissues ». Clinics in Laboratory Medicine 29, n^o 1 (mars 2009) : 101–13. http://dx.doi.org/10.1016/j.cll.2009.01.006.

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Gámez‐Pozo, Angelo, Iker Sánchez‐Navarro, Nuria Ibarz Ferrer, Fernando García Martínez, Keith Ashman et Juan Ángel Fresno Vara. « High‐Throughput Phosphoproteomics from Formalin‐Fixed, Paraffin‐Embedded Tissues ». Current Protocols in Chemical Biology 4, n^o 2 (juin 2012) : 161–75. http://dx.doi.org/10.1002/9780470559277.ch110242.

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Farragher, Susan M., Austin Tanney, Richard D. Kennedy et D. Paul Harkin. « RNA expression analysis from formalin fixed paraffin embedded tissues ». Histochemistry and Cell Biology 130, n^o 3 (5 août 2008) : 435–45. http://dx.doi.org/10.1007/s00418-008-0479-7.

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Shi, Shan-Rong, Richard J. Cote, Lin Wu, Cheng Liu, Ram Datar, Yan Shi, Dongxin Liu, Hyoeun Lim et Clive R. Taylor. « DNA Extraction from Archival Formalin-fixed, Paraffin-embedded Tissue Sections Based on the Antigen Retrieval Principle : Heating Under the Influence of pH ». Journal of Histochemistry & ; Cytochemistry 50, n^o 8 (août 2002) : 1005–11. http://dx.doi.org/10.1177/002215540205000802.

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During the course of diagnostic surgical pathology, pathologists have established a large collection of formalin-fixed, paraffin-embedded tissues that form invaluable resources for translational studies of cancer and a variety of other diseases. Accessibility of macromolecules in the fixed tissue specimens is a critical issue as exemplified by heat-induced antigen retrieval (AR) immunohistochemical (IHC) staining. On the basis of observations that heating may also enhance in situ hybridization (ISH) and the similarity of formalin-induced chemical modifications that occur in protein and in DNA, we designed a study to examine the efficiency of DNA extraction from archival formalin-fixed, paraffin-embedded tissues using an adaptation of the basic principles of the AR technique, i.e., heating the tissue under the influence of different pH values. Archival paraffin blocks of lymph nodes, tonsil, and colon were randomly selected. Each paraffin block was prepared in 34 microtubes. For each paraffin block, one tube was used as a control sample, using a non-heating DNA extraction protocol. The other 33 tubes were tested using a heating protocol under 11 variable pH values (pH 2 to 12) under three different heating conditions (80, 100, and 120C). Evaluation of the results of DNA extraction was carried out by measuring yields by photometry and PCR amplification, as well as kinetic thermocycling (KTC)-PCR methods. In general, lower pH (acid) solutions gave inferior results to solutions at higher pH (alkaline). Heating tissues at a higher temperature and at pH 6–9 gave higher yields of DNA. There appeared to be a peak in terms of highest efficiency of extracted DNA at around pH 9. The average ratios 260:280 of extracted DNA also showed better values for samples heated at 120C. PCR products of three primers showed satisfactory results for DNA extracted from archival paraffin-embedded tissues by heating protocols at pH 6–12, with results that were comparable to the control sample subjected to the standard non-heating, enzymatic DNA extraction method. This study is the first to document the use of heating at an alkaline pH for DNA extraction from archival formalin-fixed, paraffin-embedded tissues, a recommendation based on the principles of AR for protein IHC. These findings may lead to a more effective protocol for DNA extraction from archival paraffin-embedded tissues and may also provide enhanced understanding of changes that occur during formalin-induced modification of nucleic acids.

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Azril, Kuo-Yuan Huang, Jonathan Hobley, Mehdi Rouhani, Wen-Lung Liu et Yeau-Ren Jeng. « A methodology to evaluate different histological preparations of soft tissues : Intervertebral disc tissues study ». Journal of Applied Biomaterials & ; Functional Materials 21 (janvier 2023) : 228080002311556. http://dx.doi.org/10.1177/22808000231155634.

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A tissue preparation method will inevitably alter the tissue content. This study aims to evaluate how different common sample preparation methods will affect the tissue morphology, biomechanical properties, and chemical composition of samples. The study focuses on intervertebral disc (IVD) tissue; however, it can be applied to other soft tissues. Raman spectroscopy synchronized with nanoindentation instrumentation was employed to investigate the compositional changes of IVD, specifically, nucleus pulposus (NP) and annulus fibrosus (AF), together with their biomechanical properties of IVD. These properties were examined through the following histological specimen types: fresh cryosection (control), fixed cryosection, and paraffin-embedded. The IVD tissue could be located using an optical microscope under three different preparation methods. Paraffin-embedded samples showed the most explicit details where the lamellae structure of AF could be identified. In terms of biomechanical properties, there was no significant difference between the fresh and fixed cryosection ( p > 0.05). In contrast, the fresh cryosection and paraffin-embedded samples showed a significant difference ( p < 0.05). It was also found that the tissue preparations affected the chemical content of the tissues and structure of the tissue, which are expected to contribute to biomechanical properties changes. Fresh cryosection and fixed cryosection samples are more promising to work with for biomechanical assessment in histological tissues. The findings fill essential gaps in the literature by providing valuable insight into the characteristics of IVD at the microscale. This study can also become a reference for a better approach to assessing the mechanical properties and chemical content of soft tissues at the microscale.

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Azzalini, Eros, Eleonora De Martino, Paolo Fattorini, Vincenzo Canzonieri, Giorgio Stanta et Serena Bonin. « Reliability of miRNA Analysis from Fixed and Paraffin-Embedded Tissues ». International Journal of Molecular Sciences 20, n^o 19 (27 septembre 2019) : 4819. http://dx.doi.org/10.3390/ijms20194819.

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In clinical practice, patients’ tissues are fixed and paraffin-embedded in order to enable histological diagnosis. Nowadays, those tissues are also used for molecular characterization. Formalin is the most used fixative worldwide, and Bouin’s solution in some worldwide institutions. Among molecular targets, micro RNAs (miRNAs), the single-stranded non-coding RNAs comprised of 18 to 24 nucleotides, have been demonstrated to be resistant to fixation and paraffin-embedding processes, with consequent possible application in clinical practice. In the present study, let-7e-5p, miR-423-3p, miR-92a-1-5p, miR-30d-5p, miR-155-5p, miR-200a-3p, and miR-429 were investigated in formalin and matched Bouin’s solution-fixed tissues of high grade serous ovarian cancers by means of real-time and droplet digital PCR (ddPCR). Micro RNAs were detectable and analyzable in both formalin- and Bouin’s-fixed specimens, but on average, higher Ct values and lower copies/µL were found in Bouin’s-fixed samples. Data from formalin-fixed samples correlated significantly for most targets with Bouin’s ones, except for let-7e-5p and miR-155-5p. This study shows that miRNAs are analyzable in both formalin- and Bouin’s-fixed specimens, with the possibility, after proper data normalization, to compare miRNA-based data from formalin-fixed samples to those of Bouin’s-fixed ones.

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Obi, Ekenedirichukwu N., Daniel A. Tellock, Gabriel J. Thomas et Timothy D. Veenstra. « Biomarker Analysis of Formalin-Fixed Paraffin-Embedded Clinical Tissues Using Proteomics ». Biomolecules 13, n^o 1 (3 janvier 2023) : 96. http://dx.doi.org/10.3390/biom13010096.

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The relatively recent developments in mass spectrometry (MS) have provided novel opportunities for this technology to impact modern medicine. One of those opportunities is in biomarker discovery and diagnostics. Key developments in sample preparation have enabled a greater range of clinical samples to be characterized at a deeper level using MS. While most of these developments have focused on blood, tissues have also been an important resource. Fresh tissues, however, are difficult to obtain for research purposes and require significant resources for long-term storage. There are millions of archived formalin-fixed paraffin-embedded (FFPE) tissues within pathology departments worldwide representing every possible tissue type including tumors that are rare or very small. Owing to the chemical technique used to preserve FFPE tissues, they were considered intractable to many newer proteomics techniques and primarily only useful for immunohistochemistry. In the past couple of decades, however, researchers have been able to develop methods to extract proteins from FFPE tissues in a form making them analyzable using state-of-the-art technologies such as MS and protein arrays. This review will discuss the history of these developments and provide examples of how they are currently being used to identify biomarkers and diagnose diseases such as cancer.

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Shetye, J. D., A. Scheynius, H. T. Mellstedt et P. Biberfeld. « Retrieval of leukocyte antigens in paraffin-embedded rat tissues. » Journal of Histochemistry & ; Cytochemistry 44, n^o 7 (juillet 1996) : 767–76. http://dx.doi.org/10.1177/44.7.8675998.

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We compared the immunohistochemical reactivity of various rat leukocyte antigens in frozen and paraffin-embedded thymus, spleen, abdominal lymph node, liver, and brain tissues of healthy Sprague-Dawley rats, fixed in various fixatives. Immune reactivity after fixation in Methacarn was superior to that of 4% neutral buffered formalin, a mercury-based fixative (B-5), or Carnoy. Microwave (MW) antigen retrieval (AR) enhanced antigen reactivity. Ten of the 11 leukocyte antigens studied could be retrieved in Methacarn-fixed, paraffin-embedded sections with a reactivity comparable to that obtained on frozen sections.

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Rigsby, W. E., D. M. Hinton, V. J. Hurst et P. C. McCaskey. « Discovery of intracellular 2,8 dihydroxyadenine crystals in bovine lymphatic and hepatic cells ». Proceedings, annual meeting, Electron Microscopy Society of America 45 (août 1987) : 906–7. http://dx.doi.org/10.1017/s042482010012881x.

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Crystalline intracellular inclusions are rarely seen in mammalian tissues and are often difficult to positively identify. Lymph node and liver tissue samples were obtained from two cows which had been rejected at the slaughter house due to the abnormal appearance of these organs in the animals. The samples were fixed in formaldehyde and some of the fixed material was embedded in paraffin. Examination of the paraffin sections with polarized light microscopy revealed the presence of numerous crystals in both hepatic and lymph tissue sections. Tissue sections were then deparaffinized in xylene, mounted, carbon coated, and examined in a Phillips 505T SEM equipped with a Tracor Northern X-ray Energy Dispersive Spectroscopy (EDS) system. Crystals were obscured by cellular components and membranes so that EDS spectra were only obtainable from whole cells. Tissue samples which had been fixed but not paraffin-embedded were dehydrated, embedded in Spurrs plastic, and sectioned.

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Shoup, David I., David E. Swayne, Daral J. Jackwood et Linda J. Saif. « Immunohistochemistry of Transmissible Gastroenteritis Virus Antigens in Fixed Paraffin-Embedded Tissues ». Journal of Veterinary Diagnostic Investigation 8, n^o 2 (avril 1996) : 161–67. http://dx.doi.org/10.1177/104063879600800204.

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An immunohistochemistry technique was developed using fixed tissues to study the presence and location of transmissible gastroenteritis virus (TGEV) antigens in situ. Experimentally infected gnotobiotic and conventional pigs as well as pigs with natural TGEV infection were examined. The staining technique was based on detection of the major structural protein of TGEV, the nucleocapsid, by using a pool of 3 monoclonal antibodies. Formalin and periodate-lysine-paraformaldehyde (PLP)-fixed intestinal tissues from a gnotobiotic pig inoculated with virulent TGEV were used to determine optimal antibody concentrations and incubation times. The intestinal tissues remained in their respective fixatives for 6 months, and serial sections were removed at sequential times and embedded in paraffin blocks. PLP and 10% neutral buffered formalin were acceptable fixatives and preserved TGEV nucleocapsid antigenicity for up to 6 months. Formalin, in comparison with PLP as a fixative, was better for preserving original tissue morphology and provided better antigen detection. Conventional crossbred pigs were inoculated with virulent TGEV, and animals were euthanized on various postexposure days. Intestinal tissues were positive for TGEV nucleocapsid antigens on postexposure days 2, 4, and 8. The immunohistochemistry technique detected TGEV antigen in stored paraffin-embedded tissues from 14 naturally infected pigs previously confirmed as positive for TGEV using a direct immunofluorescence assay on intestinal mucosal smears, whereas 9 naturally infected pigs confirmed negative for TGEV antigen by the same immunofluorescence assay showed no staining consistent with the presence of TGEV antigen. Immunohistochemistry provides a method to detect TGEV and possibly other closely related coronaviruses such as porcine respiratory coronavirus in situ. A diagnostic test using the same fixed tissues processed for histopathology provides veterinary practitioners an alternative to delivering live pigs or refrigerated fresh intestinal samples containing infectious virus to a diagnostic laboratory. Investigators can utilize this technique to retrospectively screen fixed tissues for TGEV antigen.

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Dear, Jonathan D., Jane E. Sykes et Danika L. Bannasch. « Quality of DNA extracted from formalin-fixed, paraffin-embedded canine tissues ». Journal of Veterinary Diagnostic Investigation 32, n^o 4 (9 juin 2020) : 556–59. http://dx.doi.org/10.1177/1040638720929637.

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Veterinary pathology tissue banks are valuable resources for genetic studies. However, limited data exist as to whether quality DNA can be extracted from these tissues for use in canine genotyping studies. We extracted DNA from 44 formalin-fixed, paraffin-embedded (FFPE) tissue blocks from dogs; 9 of these dogs had DNA available from whole blood samples that had been banked. We genotyped DNA from 30 of 44 tissue blocks and 9 whole blood samples on the Illumina CanineHD BeadChip; DNA quality was insufficient in 14 of 44 samples from tissue blocks. There was significant correlation between the 260/280 ratio and single-nucleotide variation (SNV) call rate ( p = 0.0276; r2 = 0.162); 23 of 30 samples from FFPE were genotyped with > 65% call rates. Median pairwise identical-by-state (IBS) analysis was 0.99 in 8 pairs of dogs with call rates > 65%. Neither age of tissue block nor specific tissue types were associated with significant differences in DNA concentration, 260/280 ratio, or SNV call rate. DNA extracted from tissue blocks can have variable quality, although comparable levels of homozygosity suggest that extracts from FFPE with call rates > 65% might provide similar results to samples from whole blood when analyzed on the Illumina CanineHD BeadChip.

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Fowler, Carol B., Robert E. Cunningham, Timothy J. O'Leary et Jeffrey T. Mason. « ‘Tissue surrogates’ as a model for archival formalin-fixed paraffin-embedded tissues ». Laboratory Investigation 87, n^o 8 (28 mai 2007) : 836–46. http://dx.doi.org/10.1038/labinvest.3700596.

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Martin, Rodger K., Kelly T. Archer et R. Michael Tuttle. « Detection of ptc in Archival Formalin-Fixed, Paraffin-Embedded Tissues ». Diagnostic Molecular Pathology 3, n^o 4 (décembre 1994) : 233–39. http://dx.doi.org/10.1097/00019606-199412000-00004.

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Lachman, Mary F., Richard W. Cartun, Carl A. Pedersen et Solon R. Cole. « Immunocytochemical Identification of Pneumocystis cariniiin Formalin-Fixed, paraffin-Embedded Tissues ». Laboratory Medicine 21, n^o 12 (1 décembre 1990) : 808–10. http://dx.doi.org/10.1093/labmed/21.12.808.

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Giusti, Laura, Cristina Angeloni et Antonio Lucacchini. « Update on proteomic studies of formalin-fixed paraffin-embedded tissues ». Expert Review of Proteomics 16, n^o 6 (16 mai 2019) : 513–20. http://dx.doi.org/10.1080/14789450.2019.1615452.

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Alkassimi, Dr Fatma, Dr Saeed Ur Rahman et Dr Praveen Arany. « OPTIMIZATION OF DIAGNOSTIC IMMUNOHISTOCHEMISTRY OF FORMALIN-fiXED, PARAffiN-EMBEDDED TISSUES ». Oral Surgery, Oral Medicine, Oral Pathology and Oral Radiology 128, n^o 1 (juillet 2019) : e38-e39. http://dx.doi.org/10.1016/j.oooo.2019.02.071.

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Ikegaya, Hiroshi, Hirotaro Iwase, Huai-Ying Zheng, Makoto Nakajima, Koichi Sakurada, Takehiko Takatori, Masashi Fukayama, Tadaichi Kitamura et Yoshiaki Yogo. « JC virus genotyping using formalin-fixed, paraffin-embedded renal tissues ». Journal of Virological Methods 126, n^o 1-2 (juin 2005) : 37–43. http://dx.doi.org/10.1016/j.jviromet.2005.01.019.

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Bodewes, Rogier, Peter R. W. A. van Run, Anita C. Schürch, Marion P. G. Koopmans, Albert D. M. E. Osterhaus, Wolfgang Baumgärtner, Thijs Kuiken et Saskia L. Smits. « Virus characterization and discovery in formalin-fixed paraffin-embedded tissues ». Journal of Virological Methods 214 (mars 2015) : 54–59. http://dx.doi.org/10.1016/j.jviromet.2015.02.002.

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Cattoretti, Giorgio, et Albert J. H. Suurmeijer. « Antigen Unmasking on Formalin-Fixed Paraffin-Embedded Tissues Using Microwaves ». Advances in Anatomic Pathology 2, n^o 1 (janvier 1995) : 2–9. http://dx.doi.org/10.1097/00125480-199501000-00002.

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Ahram, Mamoun, Michael J. Flaig, John W. Gillespie, Paul H. Duray, W. Marston Linehan, David K. Ornstein, Shulan Niu, Yingming Zhao, Emanuel F. Petricoin et Michael R. Emmert-Buck. « Evaluation of ethanol-fixed, paraffin-embedded tissues for proteomic applications ». PROTEOMICS 3, n^o 4 (avril 2003) : 413–21. http://dx.doi.org/10.1002/pmic.200390056.

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Magdeldin, Sameh, et Tadashi Yamamoto. « Toward deciphering proteomes of formalin-fixed paraffin-embedded (FFPE) tissues ». PROTEOMICS 12, n^o 7 (avril 2012) : 1045–58. http://dx.doi.org/10.1002/pmic.201100550.

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Kothmaier, Hannelore, Daniela Rohrer, Elvira Stacher, Franz Quehenberger, Karl-Friedrich Becker et Helmut H. Popper. « Comparison of Formalin-Free Tissue Fixatives : A Proteomic Study Testing Their Application for Routine Pathology and Research ». Archives of Pathology & ; Laboratory Medicine 135, n^o 6 (1 juin 2011) : 744–52. http://dx.doi.org/10.5858/2009-0676-oa.1.

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Abstract Context.—Formalin-fixed, paraffin-embedded tissue is the routine processing method for diagnostics practiced in pathology departments worldwide. Objective.—To determine the potential value of non–cross-linking, formalin-free tissue fixation for diagnostics in pathology and proteomic investigations. Design.—We tested 3 commercially available, formalin-free tissue fixatives—FineFIX, RCL2, and HOPE—in lung cancer specimens from 10 patients. The fixatives were evaluated for their effects on tissue morphology, protein recovery, and immunoreactivity for a selected panel of proteins differently expressed in lung cancer, using immunohistochemistry and Western blotting. Results.—Tumor-cell analysis with hematoxylin-eosin worked equally well for all tested fixatives when compared with the standard formalin-fixed, paraffin-embedded procedure. Movat pentachrome stains showed comparable results for the different matrices and cellular proteins analyzed. The RCL2 (P = .01) and HOPE fixatives (P = .03) improved protein recovery when compared with formalin-fixed, paraffin-embedded or frozen tissues. Our data clearly show that the fixatives evaluated influenced immunoreactivity to matched, formalin-fixed, paraffin-embedded lung cancer tissue. In particular, membrane-bound proteins, such as epidermal growth factor receptor EGFR, can be detected more efficiently by immunohistochemistry and Western blotting. Conclusion.—We have demonstrated that formalin-free fixatives have the potential in routine pathology and research to replace formalin in histomorphology and protein preservation.

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Mason, Gillian I., et John B. Matthews. « A Comparison of Lectin Binding to Frozen, Formalin Fixed Paraffin Embedded, and Acid Decalcified Paraffin Embedded Tissues ». Journal of Histotechnology 11, n^o 4 (décembre 1988) : 223–29. http://dx.doi.org/10.1179/his.1988.11.4.223.

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Sequeiros, Tamara, Marta García, Melania Montes, Mireia Oliván, Marina Rigau, Eva Colás, Inés de Torres, Juan Morote, Jaume Reventós et Andreas Doll. « Molecular Markers for Prostate Cancer in Formalin-Fixed Paraffin-Embedded Tissues ». BioMed Research International 2013 (2013) : 1–15. http://dx.doi.org/10.1155/2013/283635.

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Prostate cancer (PCa) is the most frequently diagnosed type of cancer in developed countries. The decisive method of diagnosis is based on the results of biopsies, morphologically evaluated to determine the presence or absence of cancer. Although this approach leads to a confident diagnosis in most cases, it can be improved by using the molecular markers present in the tissue. Both miRNAs and proteins are considered excellent candidates for biomarkers in formalin-fixed paraffin-embedded (FFPE) tissues, due to their stability over long periods of time. In the last few years, a concerted effort has been made to develop the necessary tools for their reliable measurement in these types of samples. Furthermore, the use of these kinds of markers may also help in establishing tumor grade and aggressiveness, as well as predicting the possible outcomes in each particular case for the different treatments available. This would aid clinicians in the decision-making process. In this review, we attempt to summarize and discuss the potential use of microRNA and protein profiles in FFPE tissue samples as markers to better predict PCa diagnosis, progression, and response to therapy.

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Urieli-Shoval, S., R. L. Meek, R. H. Hanson, M. Ferguson, D. Gordon et E. P. Benditt. « Preservation of RNA for in situ hybridization : Carnoy's versus formaldehyde fixation. » Journal of Histochemistry & ; Cytochemistry 40, n^o 12 (décembre 1992) : 1879–85. http://dx.doi.org/10.1177/40.12.1280665.

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Tissues fixed with organic solvent fixatives such as Carnoy's solution are known to give poor and erratic results with in situ hybridization, whereas those fixed with paraformaldehyde produce more consistent results. To understand this difference and to improve the utility of Carnoy's-fixed tissue for in situ hybridization, we explored several parameters of RNA integrity and preservation. Carnoy's-fixed, paraffin-embedded livers and paraformaldehyde-fixed, paraffin-embedded livers of mice were compared for RNA extractability, degradation, and hybridizability. In addition, retention of RNA in tissue sections after sequential in situ hybridization treatments was compared. RNA was found to be easily extractable from Carnoy's-fixed liver and was well preserved, with only slight degradation of high molecular weight RNA. Conversely, only a small percentage of the RNA was extractable from paraformaldehyde-fixed liver unless the tissue was digested with protease. The extracted RNA was well preserved, without detectable degradation. Sections of tissue fixed in Carnoy's solution subjected to in situ hybridization retained only about 10% of their original RNA content and gave correspondingly weak in situ hybridization signals. Formaldehyde-fixed tissues retained much more of the RNA (about 45%) and produced strong in situ hybridization signals. Treatment of Carnoy's-fixed tissue sections with vaporous formaldehyde increased retention of RNA and provided in situ hybridization signals comparable with those of paraformaldehyde-fixed tissues.

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Conti, C. J., F. Larcher, J. Chesner et C. M. Aldaz. « Polyacrylamide gel electrophoresis and immunoblotting of proteins extracted from paraffin-embedded tissue sections. » Journal of Histochemistry & ; Cytochemistry 36, n^o 5 (mai 1988) : 547–50. http://dx.doi.org/10.1177/36.5.3282007.

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We show in this communication that polyacrylamide gel electrophoresis (PAGE) and immunoblotting of proteins can be performed using one to two 5-7 micron paraffin sections of tissues fixed in non-cross-linking fixatives (acetone, alcohol, or modified Carnoy's solution). Proteins for study were extracted from paraffin sections of mouse foot pad and liver. The presence of unaltered keratin polypeptides in tissues fixed with either acetone or alcohol was demonstrated in gels stained with Coomassie brilliant blue. The preservation of their antigenic determinants was demonstrated with immunoblotting. Furthermore, the immunoreactivity of soluble proteins, such as albumin, remained unaltered in immunoblots obtained from paraffin-embedded mouse liver sections. These data indicate that tissues embedded and stored in paraffin are useful for the above-mentioned biochemical and immunological studies and may therefore be an important technique for diagnostic pathology or retrospective studies.

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Ben-Ezra, J., D. A. Johnson, J. Rossi, N. Cook et A. Wu. « Effect of fixation on the amplification of nucleic acids from paraffin-embedded material by the polymerase chain reaction. » Journal of Histochemistry & ; Cytochemistry 39, n^o 3 (mars 1991) : 351–54. http://dx.doi.org/10.1177/39.3.1704393.

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Amplification of nucleic acids from paraffin-embedded material by the polymerase chain reaction (PCR) is increasingly being used to detect viral genomes and oncogene mutations. To determine the effect of fixation on the preservation of the nucleic acids, we fixed two randomly chosen fresh pathology specimens in formalin, B-5, Bouin's, Zenker's, ethanol, and Omnifix for 6, 24, 48, 72, and 168 hr (1 week), and then embedded the tissue in paraffin. Oligonucleotide primers specific for the cytoplasmic-beta-actin gene were chosen to span an intron such that amplification yielded a product of 250 BP for DNA and 154 BP for RNA. A single 6-microns section was cut from each paraffin block, deparaffinized, and then subjected to 30 rounds of amplification for either DNA or RNA. On amplifying DNA, consistent product was seen in the ethanol and Omnifix specimens up to 72 hr of fixation time, whereas variable product was seen with formalin or Zenker's fixation; all specimens fixed in Bouin's or B-5 were negative. On amplifying RNA, a product could be detected even after 1 week of fixation in ethanol or Omnifix, and after 48 hr in the formalin-fixed tissue. The Zenker's-fixed tissues gave variable results, and the Bouin's and B-5 tissues gave consistent results only after 6 hr of fixation. We therefore conclude that choice of fixative and fixation time are critical factors influencing the outcome of PCR amplification of nucleic acids from paraffin-embedded material.

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de las Mulas, Juana Martín, Eduardo Ruiz-Villamor, Sergio Donoso, Manuel Quezada, Claudio Lecocq et Miguel Angel Sierra. « Immunohistochemical Detection of Hog Cholera Viral Glycoprotein 55 in Paraffin-Embedded Tissues ». Journal of Veterinary Diagnostic Investigation 9, n^o 1 (janvier 1997) : 10–16. http://dx.doi.org/10.1177/104063879700900103.

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Formalin-fixed, paraffin embedded tissues obtained from 40 pigs inoculated with a field isolate of hog cholera virus were examined for the presence of Gp55, a major structural protein of the virus envelope, using a monoclonal antibody-based immunohistochemical test with the avidin-biotin-peroxidase complex method. Immunoreactivity was detected in hog cholera virus-infected tissues but not in control pigs tissues, African swine fever virus-infected tissues, or bovine viral diarrhea virus-infected porcine or bovine tissues. The first positive reactions were seen in lymphatic tissues, digestive tract and skin on postinoculation day (pid) 4, respiratory and urinary tissues on pid 5, nervous tissues on pid 6, and endocrine tissues on pid 7. These staining reactions persisted until the last observation on pid 18. Hog cholera virus antigen was not detected in heart tissue at any time. The highest levels of antigen detection were found in tonsils, spleen, and pancreas, although the esophageal mucosa and skin epithelial cells were also intensely and widely stained. The cellular staining pattern of Gp55 had a ubiquitous distribution. It was found in epithelial cells, macrophages and circulating monocytes, endothelial cells, lymphoid cells, and glial cells. The results showed a high specificity and high sensitivity for detecting hog cholera Gp55 in formalin-fixed, paraffin embedded tissue samples. This method allows precise association of Gp55 with specific cells, tissues, and histologic lesions, making the technique suitable for use in routine diagnosis of hog cholera.

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Bolognesi, Maddalena M., Francesco Mascadri, Laura Furia, Mario Faretta, Francesca M. Bosisio et Giorgio Cattoretti. « Antibodies validated for routinely processed tissues stain frozen sections unpredictably ». BioTechniques 70, n^o 3 (mars 2021) : 137–48. http://dx.doi.org/10.2144/btn-2020-0149.

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Background: Antibody validation for tissue staining is required for reproducibility; criteria to ensure validity have been published recently. The majority of these recommendations imply the use of routinely processed (formalin-fixed, paraffin-embedded) tissue. Materials & methods: We applied to lightly fixed frozen sections a panel of 126 antibodies validated for formalin-fixed, paraffin-embedded tissue with extended criteria. Results: Less than 30% of the antibodies performed as expected with all fixations. 35% preferred one fixation over another, 13% gave nonspecific staining and 23% did not stain at all. Conclusion: Individual antibody variability of the paratope’s fitness for the fixed antigen may be the cause. Revalidation of established antibody panels is required when they are applied to sections whose fixation and processing are different from the tissue where they were initially validated.

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Janssen, P. J., A. O. Brinkmann, W. J. Boersma et T. H. Van der Kwast. « Immunohistochemical detection of the androgen receptor with monoclonal antibody F39.4 in routinely processed, paraffin-embedded human tissues after microwave pre-treatment. » Journal of Histochemistry & ; Cytochemistry 42, n^o 8 (août 1994) : 1169–75. http://dx.doi.org/10.1177/42.8.8027537.

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We describe the immunohistochemical detection of the human androgen receptor (AR) in routinely processed, paraffin-embedded tissue with the monoclonal antibody (MAb) F39.4. Deparaffinized sections were heated in a microwave oven for antigen retrieval. A panel of human male- and female-derived tissues was investigated. We observed a nuclear staining pattern consistent with previous results on frozen sections. Moreover, we studied the possibility of detecting AR in prolonged formalin-fixed tissue and in paraffin-embedded archival material. After prolonged fixation times or long-term storage of paraffin-embedded tissue, the staining intensity for the AR did not deteriorate. Blocking experiments with the specific synthetic peptides demonstrated the specificity of this technique. We conclude that this method is specific, allows retrospective AR studies, and offers optimally preserved morphology.

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Nucci, Ricardo, Wilson Jacob-Filho, Alexandre Busse, Laura Maifrino et Romeu de Souza. « Anatomopathological Assessment of the Diaphragm in Formalin-Fixed, Paraffin-Embedded Sections ». Journal of Morphological Sciences 35, n^o 03 (septembre 2018) : 173–76. http://dx.doi.org/10.1055/s-0038-1673611.

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Introduction The analysis of frozen muscle biopsies has become a routine method in the evaluation of muscle structure in health and disease. However, the technique for frozen muscle specimens is not widely available in countries with limited medical facilities. The present study aimed to elucidate a reproducible formalin-fixed and paraffin-embedded (FFPE) method for this type of analysis in postmortem muscles. Methods Diaphragm muscle was obtained within 1 hour of sudden death. Diaphragm strips were washed in saline solution, fixed in 10% formalin, frozen at 4°C in a refrigerator, and stored for 24 hours. Then, the tissue samples were processed into paraffin-embedded blocks. Transversal sections were cut from each paraffin block and stained with hematoxylin and eosin, Picrosirius red, Verhoeff-Van Gieson, and Congo red for the qualitative analysis. Results Our analysis indicated a well-preserved muscle. Conclusion In summary, we demonstrate a simple technique for a reproducible FFPE method in postmortem muscle tissues.

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Frickmann, Hagen, Ulrike Loderstaedt, Paul Racz, Klara Tenner-Racz, Petra Eggert, Alexandra Haeupler, Ralf Bialek et Ralf Matthias Hagen. « Detection of Tropical Fungi in Formalin-Fixed, Paraffin-Embedded Tissue : Still an Indication for Microscopy in Times of Sequence-Based Diagnosis ? » BioMed Research International 2015 (2015) : 1–11. http://dx.doi.org/10.1155/2015/938721.

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Introduction. The aim of the study was the evaluation of panfungal PCR protocols with subsequent sequence analysis for the diagnostic identification of invasive mycoses in formalin-fixed, paraffin-embedded tissue samples with rare tropical mycoses.Materials and Methods. Five different previously described panfungal PCR/sequencing protocols targeting 18S and 28S ribosomal RNA gene fragments as well as internal transcribed spacer 1 and 2 fragments were evaluated with a collection of 17 formalin-fixed, paraffin-embedded tissue samples of patients with rare and/or tropical invasive mycoses, comprising chromoblastomycosis, coccidioidomycosis, cryptococcosis, histoplasmosis, mucormycosis, mycetoma/maduromycosis, and rhinosporidiosis, in a proof-of-principle analysis.Results. The primers of the panfungal PCRs readily and predominantly reacted with contaminating environmental fungi that had deposited on the paraffin blocks. Altogether three sequence results of histoplasmosis and mycetoma samples that matched the histological assessment were associated with sample age <10 years and virtually without PCR inhibition.Conclusions. The high risk of amplifying environmental contaminants severely reduces the usefulness of the assessed panfungal PCR/sequencing protocols for the identification of rare and/or tropical mycoses in stored formalin-fixed, paraffin-embedded tissues. Histological assessment remains valuable for such indications if cultural differentiation is impossible from inactivated sample material.

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Yoo, Jinyoung, Seok Jin Kang et Chang Suk Kang. « Telomerase Activity in Gastric Adenocarcinomas : Frozen Tissues Versus Methacarn-fixed Paraffin-embedded Tissues ». Cancer Research and Treatment 35, n^o 6 (31 décembre 2003) : 478–82. http://dx.doi.org/10.4143/crt.2003.35.6.478.

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Hayashi, Y., M. Koike, M. Matsutani et T. Hoshino. « Effects of fixation time and enzymatic digestion on immunohistochemical demonstration of bromodeoxyuridine in formalin-fixed, paraffin-embedded tissue. » Journal of Histochemistry & ; Cytochemistry 36, n^o 5 (mai 1988) : 511–14. http://dx.doi.org/10.1177/36.5.3282006.

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We studied the effects of fixation time and enzymatic digestion on immunohistochemical staining for bromodeoxyuridine (BUdR) in excised rat and human gastrointestinal tissues and human brain tumors which had been fixed in formalin after intravenous administration of BUdR shortly before biopsy of tissue. In formalin-fixed rat gastrointestinal tissues not treated with proteinase, the reaction products were insufficient to identify BUdR-positive cells. Results similar to those in ethanol-fixed tissue were obtained when formalin-fixed tissue sections were treated with protease, pepsin, or trypsin. The longer the material had been fixed in formalin, the longer the incubation in proteinase required to identify BUdR-labeled nuclei. The BUdR labeling indices of formalin-fixed human brain tumor specimens treated with protease were comparable to those of ethanol-fixed tissues. Sufficient BUdR staining was obtained even in tissues fixed in formalin for prolonged periods. Therefore, the BUdR labeling index can be determined retrospectively in clinical materials stored in formalin.

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Mills, Anne, Rajeev Balasubramaniam, Teri A. Longacre, Christina S. Kong et Benjamin A. Pinsky. « Laboratory-Developed L1 Sequencing and Type-Specific, Real-Time Polymerase Chain Reaction for the Detection and Typing of Human Papillomaviruses in Formalin-Fixed, Paraffin-Embedded Tissues ». Archives of Pathology & ; Laboratory Medicine 137, n^o 1 (1 janvier 2013) : 50–54. http://dx.doi.org/10.5858/arpa.2011-0392-oa.

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Context.—The detection and typing of high-risk and low-risk human papillomavirus (HPV) in archival formalin-fixed, paraffin-embedded tissues by nucleic acid amplification testing is an important adjunct to immunohistochemical staining in evaluation of squamous cell proliferations of the oropharynx, larynx, and anal canal. Objective.—To evaluate semiautomated, xylene-free extraction from formalin-fixed, paraffin-embedded tissues combined with laboratory-developed HPV L1 sequencing and type-specific HPV 6, 11, 16, and 18 real-time polymerase chain reaction for identification and typing of HPV in the clinical laboratory. Design.—We evaluated the adequacy of extraction using β-globin amplification and compared L1 sequencing and real-time polymerase chain reaction methods for typing accuracy using 68 formalin-fixed, paraffin-embedded tissues, including 56 anorectal biopsy or surgical resection specimens and 12 laryngeal papilloma specimens from patients with recurrent respiratory papillomatosis. Results.—Adequate DNA was obtained from 68 of 68 specimens analyzed and all were HPV positive. In 47 cases where L1 sequencing demonstrated that the predominant HPV type was 6, 11, 16, or 18, type-specific, real-time polymerase chain reaction provided concordant results. Sequencing revealed additional low-risk (HPV 40) and high-risk HPV types (HPV 31, 33, 56, and 58) in anorectal specimens, whereas HPV 6 or 11 were the types found in laryngeal papillomas. Conclusion.—Both L1 sequencing and type-specific, real-time polymerase chain reaction are suitable methods for routine HPV testing of formalin-fixed, paraffin-embedded tissues in a clinical laboratory setting.

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Dimaras, Pantelis, Oskan Tasinov, Desislava Ivanova, Yoana Kiselova-Kaneva, Nadezhda Stefanova, Maria Tzaneva et Diana Ivanova. « Improving gene expression analysis efficacy from formalin-fixed paraffin embedded tissues ». Folia Medica 64, n^o 4 (31 août 2022) : 602–8. http://dx.doi.org/10.3897/folmed.64.e63599.

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Introduction: Improving RNA isolation and cDNA synthesis techniques has emerged due to advancements in the knowledge of molecular basis of most diseases. This in turn increased the need of higher quantity and quality of the extracted genetic material to be used for a variety of diagnostic tests and experiments. Aim: The aim of the study was to compare three modified methods for RNA extraction from formalin-fixed paraffin embedded (FFPE) biopsied tissue and different cDNA synthesis strategies to facilitate study of gene expression. Materials and methods: Compared RNA extraction methods were: lysis buffer, phenol-based extraction, and combination of both with concomitant use of silica-based spin columns. RNA quantity and purity were estimated spectrophotometrically. Different priming strategies for cDNA synthesis were applied: oligo dT, combination of oligo dT and random hexamer, and gene specific primer. Two-step RT-qPCR of ribosomal protein L37A on preamplified and non-preamplified cDNA templates was performed. Results: The combination of lysis buffer with phenol based extraction gave higher RNA yield. By doing cDNA preamplification, the confidence of detection by qPCR was raised, and efficiency was improved. The preamplified template increased the sensitivity of analysis. Conclusions: Together, the combination of approaches improved substantially the reproducibility and validity of quantitative gene expression analyses from FFPE tissues.

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Baszler, Timothy V., Lawrence J. C. Gay, Maureen T. Long et Bruce A. Mathison. « Detection by PCR of Neospora caninum in Fetal Tissues from Spontaneous Bovine Abortions ». Journal of Clinical Microbiology 37, n^o 12 (1999) : 4059–64. http://dx.doi.org/10.1128/jcm.37.12.4059-4064.1999.

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The routine diagnosis of Neospora caninum abortion is based upon histopathologic changes in fetal tissues and identification of tissue parasites by immunohistochemistry. Confirmation of N. caninum infection by immunohistochemistry has low sensitivity. In the present study, we examined the utility of PCR in detecting N. caninum infection in fetal tissues from spontaneous bovine abortion. DNA was obtained from fresh and formalin-fixed tissues from 61 bovine fetuses submitted for abortion diagnosis. Histopathology and immunohistochemistry determined the true status of N. caninum infection in each fetus. In formalin-fixed paraffin-embedded tissues, PCR detected N. caninum DNA in 13 of 13 true-positive fetuses (100%) and in 1 of 16 true-negative fetuses (6%). In fresh or frozen tissues, PCR detected N. caninum DNA in 10 of 13 true-positive fetuses (77%) and 0 of 11 true-negative fetuses (0%). PCR also detected N. caninumDNA in 6 of 8 fetuses that had typical lesions of N. caninum but were immunohistochemistry negative, indicating a higher sensitivity of PCR in comparison to that of immunohistochemistry. N. caninum DNA was amplified most consistently from brain tissue. PCR detection of N. caninumDNA in formalin-fixed, paraffin-embedded tissues was superior to that in fresh tissues, presumably because of the increased accuracy of sample selection inherent in histologic specimens.

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Weke, Kenneth, Sachin Kote, Jakub Faktor, Sofian Al Shboul, Naomi Uwugiaren, Paul M. Brennan, David R. Goodlett, Ted R. Hupp et Irena Dapic. « DIA-MS proteome analysis of formalin-fixed paraffin-embedded glioblastoma tissues ». Analytica Chimica Acta 1204 (avril 2022) : 339695. http://dx.doi.org/10.1016/j.aca.2022.339695.

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Tian, Yuan, Kay Gurley, Danni L. Meany, Christopher J. Kemp et Hui Zhang. « N-Linked Glycoproteomic Analysis of Formalin-Fixed and Paraffin-Embedded Tissues ». Journal of Proteome Research 8, n^o 4 (3 avril 2009) : 1657–62. http://dx.doi.org/10.1021/pr800952h.

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Sikora, M. J., J. N. Thibert, J. Salter, M. Dowsett, M. D. Johnson et J. M. Rae. « High-efficiency genotype analysis from formalin-fixed, paraffin-embedded tumor tissues ». Pharmacogenomics Journal 11, n^o 5 (15 juin 2010) : 348–58. http://dx.doi.org/10.1038/tpj.2010.50.

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Wu, Lin, Nancy Patten, Carl T. Yamashiro et Buena Chui. « Extraction and Amplification of DNA From Formalin-Fixed, Paraffin-Embedded Tissues ». Applied Immunohistochemistry & ; Molecular Morphology 10, n^o 3 (septembre 2002) : 269–74. http://dx.doi.org/10.1097/00129039-200209000-00015.

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Chen, Jun, Gerald E. Byrne et Izidore S. Lossos. « Optimization of RNA Extraction From Formalin-fixed, Paraffin-embedded Lymphoid Tissues ». Diagnostic Molecular Pathology 16, n^o 2 (juin 2007) : 61–72. http://dx.doi.org/10.1097/pdm.0b013e31802f0804.

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Karatas, S., J. Mikalsen, T. M. Steinum, T. Taksdal, M. Bordevik et D. J. Colquhoun. « Real time PCR detection ofPiscirickettsia salmonisfrom formalin-fixed paraffin-embedded tissues ». Journal of Fish Diseases 31, n^o 10 (octobre 2008) : 747–53. http://dx.doi.org/10.1111/j.1365-2761.2008.00948.x.

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