Detection of Immune System Cells in Paraffin Wax-embedded Ovine Tissues

González, Lorenzo; Anderson, I.E.; Deane, David; Summers, Christina; Buxton, D.

doi:10.1053/jcpa.2001.0475

Cited by 83 publications

(45 citation statements)

References 17 publications

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“…Recently, Beckstead (Beckstead, 1994) presented a zinc-based fixative without cross-linking or coagulating agents but with retained morphologic qualities as well as preserved immunoreactivity. These results have also been confirmed by other investigators (Gonzalez et al, 2001;Johansson et al, 2000;Nitta et al, 1997;Wang et al, 1996).…”

supporting

confidence: 87%

“…The use of ZBF has proven advantageous in immunohistochemical staining of paraffin-embedded tissue sections (Beckstead, 1994;Gonzalez et al, 2001;Nitta et al, 1997;Wang et al, 1996). Furthermore, according to Beckstead (Beckstead, 1994) no apparent loss of antigen was observed in paraffin blocks stored for over 3 years at room temperature.…”

Section: Zinc-based Fixative In Histoprocessing Ofmentioning

confidence: 99%

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Zinc-Based Fixative Improves Preservation of Genomic DNA and Proteins in Histoprocessing of Human Tissues

Wester

Asplund

Bäckvall

et al. 2003

Laboratory Investigation

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SUMMARY:Advantageous preservation of histology and detailed cellular morphology has rendered neutral buffered formalin (NBF) the most widely used fixative in clinical pathology. Despite excellent morphology for routine diagnostics, a major drawback of NBF fixation is its detrimental effect on DNA and RNA quality. In addition to complicating analysis of genes and transcripts in complex tissues, NBF denatures proteins and thereby hampers immunohistochemical visualization of certain antigens. In the present study, we evaluated a zinc-based fixative (ZBF) regarding its effects on tissue morphology, quality of genomic DNA, and preservation of protein immunoreactivity in a broad spectrum of tissues. Four different modes of fixation were analyzed: ZBF-paraffin embedding, NBF-paraffin embedding, ZBF-fixation prior to snap-freezing, and immediate snap-freezing. Laserassisted microdissection, allowing retrieval of a defined number of cells for PCR, was used to study DNA quality. Genomic DNA was analyzed using primers for ␤2-microglobulin and the transferrin receptor. Immunohistochemistry was performed using nine antibodies. Tissue microarray blocks were used for analysis of morphology and immunoreactivity. Only slight impairment of morphologic qualities was found after ZBF-paraffin embedding, whereas ZBF prior to freezing resulted in a more crisp morphology compared with routine cryosections. A significantly higher DNA yield was observed in samples isolated from ZBF-paraffin-embedded tissues compared with NBF-paraffin-embedded tissues. Both yield and quality of DNA was comparable in frozen tissues irrespective to prior ZBF fixation. Immunoreactivity in paraffin-embedded tissue was superior in ZBF-fixated tissue compared with NBF-fixated for a majority of tested antibodies. Furthermore, for seven out of nine antibodies, antigen retrieval pretreatment proved unnecessary in ZBF-fixated tissue. Thus, despite a slight impairment of morphology, ZBF preserves protein structures well. We conclude that ZBF is superior to NBF for analysis of DNA and protein expression. Fixation of tissues in ZBF may also be an alternative strategy to freeze storage of tissue specimens, eg, in future bio-banks. (Lab Invest 2003, 83:889 -899).

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supporting

confidence: 87%

Section: Zinc-based Fixative In Histoprocessing Ofmentioning

confidence: 99%

Zinc-Based Fixative Improves Preservation of Genomic DNA and Proteins in Histoprocessing of Human Tissues

Wester

Asplund

Bäckvall

et al. 2003

Laboratory Investigation

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“…Embryos (E10.5–E15.5) and dissected kidneys (E17.5, neonates, and mice older than 8 weeks) were frozen in liquid nitrogen. Embryos E12.5 and kidneys from mice older than 8 weeks were fixed in zinc solution [15] and embedded in paraffin. All embryos (E10.5–E17.5) and neonates (P1–P7) were typed for the presence of hEpCAM by immunohistochemistry.…”

Section: Methodsmentioning

confidence: 99%

Spatial and Temporal Expression Patterns of the Epithelial Cell Adhesion Molecule (EpCAM/EGP-2) in Developing and Adult Kidneys

Trzpis¹,

Popa²,

McLaughlin³

et al. 2007

Nephron Exp Nephrol

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Background: The epithelial cell adhesion molecule (EpCAM) is expressed by most epithelia and is involved in processes fundamental for morphogenesis, including cell-cell adhesion, proliferation, differentiation, and migration. Previously, a role for EpCAM in pancreatic morphogenesis was confirmed in vitro. Furthermore, changes in the EpCAM expression pattern were found in developing lung and thymus and in the regenerating liver. Therefore, EpCAM was proposed to be a morphoregulatory molecule. Methods: Using immunohistochemistry, the expression pattern of human and murine homologues of EpCAM was characterized in adult and embryonic kidneys from humans and human-EpCAM (hEpCAM)-transgenic mice. Results: EpCAM expression was found in the ureteric bud throughout nephrogenesis. EpCAM was not expressed in the metanephric mesenchyme. In comma- and S-shaped bodies, both metanephric mesenchyme derived structures, EpCAM expression appeared by E13.5. In adult kidneys, most epithelia expressed varying levels of EpCAM, as confirmed by double staining for human EpCAM and segment-specific nephron markers. Podocytes were EpCAM negative. At the cellular level, the EpCAM expression shifted from apical in embryonic to basolateral in adult kidneys. Conclusions: The spatiotemporal expression pattern of EpCAM changes during nephrogenesis. In the adult kidney, the expression varies markedly along the nephron. These data provide a basis for further studies on EpCAM in developing and adult kidneys.

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“…In order to check the purity of the epithelial cell population isolated from the gastric mucosa, gastric samples were fixed in zinc salt solutions at slaughter [8] for embedding and sectioning. Staining of ovine epithelial cells was achieved with mouse antibodies to pancytokeratin (1:2 000, Sigma; clone PCK-26; broad spectrum antibody; reacts specifically with a variety of normal, reactive, and neoplastic epithelial tissues).…”

Section: Immunohistochemistrymentioning

confidence: 99%

Proteomic approach to identify candidate effector molecules during the in vitro immune exclusion of infectiveTeladorsagia circumcinctain the abomasum of sheep

Athanasiadou

Pemberton²,

Jackson³

et al. 2008

Vet. Res.

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-In the present study we have employed an in vitro organ challenge model to study the post-challenge responses in parasite naïve and immune gastric tissue of sheep, in an attempt to identify the host derived factors involved in immune exclusion of Teladorsagia circumcincta larvae. Proteins present in the epithelial cells and mucus from ovine abomasa following parasite challenge in previously naïve and immune animals were analysed through Matrix Assisted Laser Desorption/Ionization-Time of Flight (MALDI-Tof)-MS and shotgun proteomics. MALDI-ToF analysis of epithelial cell lysates revealed that a number of proteins identified were differentially expressed in naïve and immune cells. These included intelectin and lysozymes, which were present at higher levels in epithelial cell lysates derived from immune samples. A large number of proteins were identified in the mucosal wash from immune tissue which were not present in the mucosal wash of the naïve tissue. Some of these proteins were present in washes of immune tissue prior to the parasite challenge including immunoglobulin A, galectin 14 and 15 and sheep mast cell protease 1. However, other proteins, such as calcium activated chloride channel and intelectin were only detected in the washings from the challenged tissue. The latter may be related to an enhanced mucus release, which may result in entrapment of infective larvae and thus reduced establishment in tissue that has been previously challenged with the parasite. In conclusion, several proteins have been identified which may be involved, either directly or indirectly, in the exclusion and immune elimination of incoming infective larvae. In the present study, the usefulness of the in vitro model has been confirmed, and the global proteomic approach has identified proteins that had not previously been associated with parasite exclusion from abomasal mucosa, such as the calcium activated chloride channel.immunity / mucus / proteomics / sheep / Teladorsagia circumcincta

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Detection of Immune System Cells in Paraffin Wax-embedded Ovine Tissues

Cited by 83 publications

References 17 publications

Zinc-Based Fixative Improves Preservation of Genomic DNA and Proteins in Histoprocessing of Human Tissues

Zinc-Based Fixative Improves Preservation of Genomic DNA and Proteins in Histoprocessing of Human Tissues

Spatial and Temporal Expression Patterns of the Epithelial Cell Adhesion Molecule (EpCAM/EGP-2) in Developing and Adult Kidneys

Proteomic approach to identify candidate effector molecules during the in vitro immune exclusion of infectiveTeladorsagia circumcinctain the abomasum of sheep

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