scholarly journals NeuN: a useful neuronal marker for diagnostic histopathology.

1996 ◽  
Vol 44 (10) ◽  
pp. 1167-1171 ◽  
Author(s):  
H K Wolf ◽  
R Buslei ◽  
R Schmidt-Kastner ◽  
P K Schmidt-Kastner ◽  
T Pietsch ◽  
...  
Keyword(s):  
Neuronal Cell ◽  
Cell Types ◽  
Specific Protein ◽  
Antigen Retrieval ◽  
Neuronal Marker ◽  
Formalin Fixed Paraffin ◽  
Wide Range ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed ◽  
Diagnostic Histopathology

The monoclonal antibody A60 specifically recognizes the DNA-binding, neuron-specific protein NeuN, which is present in most neuronal cell types of vertebrates. In this study we demonstrate the potential use of NeuN as a diagnostic neuronal marker using a wide range of formalin-fixed, paraffin-embedded human surgical and autopsy specimens from the central and peripheral nervous system. After microwave antigen retrieval, almost all neuronal populations revealed strong immunoreactivity for NeuN in nuclei, perikarya, and some proximal neuronal processes, whereas more distal axon cylinders and dendritic ramifications were not stained. The stain greatly enhanced the gray matter architecture. NeuN immunoreactivity was not detected in Purkinje cells, most neurons of the internal nuclear layer of the retina, and in sympathetic chain ganglia. We examined nine gangliogliomas and 14 dysembryoplastic neuroepithelial tumors, one ganglioneuroma, and one dysplastic cerebellar gangliocytoma. The neuronal component of all of these lesions showed marked immunoreactivity for NeuN. In addition, NeuN immunoreactivity was focally seen in one of seven medulloblastomas with prominent neuronal differentiation. There was no staining of non-neuronal structures. The results indicate that NeuN immunoreactivity is a sensitive and specific neuronal marker in formalin-fixed, paraffin-embedded tissues, and may be useful in diagnostic histopathology.

Immunohistochemical Detection of Canine Adenovirus in Paraffin Sections of Liver

1986 ◽  
Vol 23 (4) ◽  
pp. 478-484 ◽  
Author(s):  
P. M. Rakich ◽  
K. W. Prasse ◽  
P. D. Lukert ◽  
L. M. Cornelius
Keyword(s):  
Immunohistochemical Detection ◽  
Hepatic Inflammation ◽  
Paraffin Sections ◽  
Inflammatory Lesions ◽  
Long Term Stability ◽  
Formalin Fixed Paraffin ◽  
Wide Range ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

An avidin-biotin immunoperoxidase procedure was optimized for detection of canine adenoviral antigens in formalin-fixed, paraffin-embedded liver. Long-term stability of viral antigen was shown by successful demonstration of virus in liver tissue preserved up to six years from dogs with infectious canine hepatitis. This immunohistochemical stain was applied to sections from livers with a wide range of inflammatory lesions. Examination of sections from 53 dogs yielded five livers with small amounts of adenovirus. An additional virus-positive liver was identified from a dog with no hepatic inflammation. Although a cause and effect relationship remains to be determined, these findings suggest a possible connection between canine adenovirus and spontaneous chronic hepatitis.

scholarly journals Immunohistochemistry for the detection of neural and inflammatory cells in equine brain tissue

PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e1601 ◽  
Author(s):  
Gretchen H. Delcambre ◽  
Junjie Liu ◽  
Jenna M. Herrington ◽  
Kelsey Vallario ◽  
Maureen T. Long
Keyword(s):  
Polyclonal Antibodies ◽  
Inflammatory Cells ◽  
Specific Protein ◽  
Proteinase K ◽  
Antigen Retrieval ◽  
Phenotypic Characterization ◽  
Buffer Solution ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Species Specific

Phenotypic characterization of cellular responses in equine infectious encephalitides has had limited description of both peripheral and resident cell populations in central nervous system (CNS) tissues due to limited species-specific reagents that react with formalin-fixed, paraffin embedded tissue (FFPE). This study identified a set of antibodies for investigating the immunopathology of infectious CNS diseases in horses. Multiple commercially available staining reagents and antibodies derived from antigens of various species for manual immunohistochemistry (IHC) were screened. Several techniques and reagents for heat-induced antigen retrieval, non-specific protein blocking, endogenous peroxidase blocking, and visualization-detection systems were tested during IHC protocol development. Boiling of slides in a low pH, citrate-based buffer solution in a double-boiler system was most consistent for epitope retrieval. Pressure-cooking, microwaving, high pH buffers, and proteinase K solutions often resulted in tissue disruption or no reactivity. Optimal blocking reagents and concentrations of each working antibody were determined. Ultimately, a set of monoclonal (mAb) and polyclonal antibodies (pAb) were identified for CD3+(pAb A0452, Dako) T-lymphocytes, CD79αcy+B-lymphocytes (mAb HM57, Dako), macrophages (mAb MAC387, Leica), NF-H+neurons (mAb NAP4, EnCor Biotechnology), microglia/macrophage (pAb Iba-1, Wako), and GFAP+astrocytes (mAb 5C10, EnCor Biotechnology). In paraffin embedded tissues, mAbs and pAbs derived from human and swine antigens were very successful at binding equine tissue targets. Individual, optimized protocols are provided for each positively reactive antibody for analyzing equine neuroinflammatory disease histopathology.

scholarly journals ­Immunohistochemistry for the detection of neural and inflammatory cells in equine brain tissue

2016 ◽  
Author(s):  
Gretchen H Delcambre ◽  
Junjie Liu ◽  
Jenna M Herrington ◽  
Kelsey Vallario ◽  
Maureen T Long
Keyword(s):  
Polyclonal Antibodies ◽  
Inflammatory Cells ◽  
Specific Protein ◽  
Proteinase K ◽  
Antigen Retrieval ◽  
Phenotypic Characterization ◽  
Buffer Solution ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Species Specific

Phenotypic characterization of cellular responses in equine infectious encephalitides has had limited description of both peripheral and resident cell populations in central nervous system (CNS) tissues due to limited species-specific reagents that react with formalin-fixed, paraffin embedded tissue (FFPE). This study identified a set of antibodies for investigating the immunopathology of infectious CNS diseases in horses. Multiple commercially available staining reagents and antibodies derived from antigens of various species for manual immunohistochemistry (IHC) were screened. Several techniques and reagents for heat-induced antigen retrieval, non-specific protein blocking, endogenous peroxidase blocking, and visualization-detection systems were tested during IHC protocol development. Boiling of slides in a low pH, citrate-based buffer solution in a double-boiler system was most consistent for epitope retrieval. Pressure-cooking, microwaving, high pH buffers, and proteinase K solutions often resulted in tissue disruption or no reactivity. Optimal blocking reagents and concentrations of each working antibody were determined. Ultimately, a set of monoclonal (mAb) and polyclonal antibodies (pAb) were identified for CD3+ (pAb A0452, Dako) T-lymphocytes, CD79αcy+ B-lymphocytes (mAb HM57, Dako), macrophages (mAb MAC387, Leica), NF-H+ neurons (mAb NAP4, EnCor Biotechnology), microglia/macrophage (pAb Iba-1, Wako), and GFAP+ astrocytes (mAb 5C10, EnCor Biotechnology). In paraffin embedded tissues, mAbs and pAbs derived from human and swine antigens were very successful at binding equine tissue targets. Individual, optimized protocols are provided for each positively reactive antibody for analyzing equine neuroinflammatory disease histopathology.

scholarly journals p53 overexpression in formalin-fixed, paraffin-embedded tissue detected by immunohistochemistry.

1992 ◽  
Vol 40 (7) ◽  
pp. 1047-1051 ◽  
Author(s):  
B J Kerns ◽  
P A Jordan ◽  
M B Moore ◽  
P A Humphrey ◽  
A Berchuck ◽  
...  
Keyword(s):  
Breast Cancers ◽  
Fresh Tissue ◽  
P53 Overexpression ◽  
P53 Expression ◽  
Ovarian Cancers ◽  
Formalin Fixed Paraffin ◽  
Wide Range ◽  
Formalin Fixed Paraffin Embedded ◽  
Fresh Frozen ◽  
Formalin Fixed

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.

scholarly journals Protein-embedding Technique: A Potential Approach to Standardization of Immunohistochemistry for Formalin-fixed, Paraffin-embedded Tissue Sections

2005 ◽  
Vol 53 (9) ◽  
pp. 1167-1170 ◽  
Author(s):  
Shan-Rong Shi ◽  
Cheng Liu ◽  
Jeanette Perez ◽  
Clive R. Taylor
Keyword(s):  
Solid Phase ◽  
Antigen Retrieval ◽  
Polymer Microsphere ◽  
Thin Sections ◽  
Tissue Sections ◽  
Embedding Technique ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Two Phases ◽  
Formalin Fixed

A serial study was performed to develop a protein-embedding technique for standardization of immunohistochemistry (IHC) on formalin-fixed, paraffin-embedded (FFPE) tissue sections. A protein carrier matrix must have two phases, a liquid phase to allow uniform mixing of a protein and a solid phase forming a ‘block’ that can be fixed and processed in the same manner as human tissue. This standardized protein block would serve as a source of thin sections for control of IHC and therefore must also withstand the boiling conditions of antigen retrieval (AR). After multiple experiments, a method was developed utilizing polymer microsphere (beads) as a support medium for protein. The method showed particular promise for quantitative IHC.

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