Hypothesis for the mechanism for heat-induced antigen retrieval occurring on fresh frozen sections without formalin-fixation in immunohistochemistry

2008 ◽  
Vol 39 (4) ◽  
pp. 389-399 ◽  
Author(s):  
Kochi Kakimoto ◽  
Susumu Takekoshi ◽  
Katsuhiro Miyajima ◽  
R. Yoshiyuki Osamura
Keyword(s):  
Formalin Fixation ◽  
Antigen Retrieval ◽  
Frozen Sections ◽  
Fresh Frozen

scholarly journals Application of Heat-induced Antigen Retrieval to Aldehyde-fixed Fresh Frozen Sections

2005 ◽  
Vol 53 (11) ◽  
pp. 1421-1432 ◽  
Author(s):  
Shuji Yamashita ◽  
Yasunori Okada
Keyword(s):  
Room Temperature ◽  
Glucocorticoid Receptors ◽  
Estrogen Receptor Α ◽  
Antigen Retrieval ◽  
Frozen Sections ◽  
New Approach ◽  
Cacodylate Buffer ◽  
Ligand Free ◽  
Fresh Frozen ◽  
Mouse Hepatocytes

We applied the heat-induced antigen retrieval (HIAR) to aldehyde-fixed fresh frozen sections based on a new approach (i.e., a rapid and complete immobilization of antigen followed by heating). Frozen sections were fixed with 10% formalin in 0.1 M cacodylate buffer (pH 7.4) containing 25 mM CaCl2 for 30 min, or with 0.5% glutaraldehyde in 0.1 M phosphate buffer (pH 7.4) for 1 min at room temperature, and then autoclaved in 20 mM Tris-HCl buffer (pH 9.0) for 10 min at 120C. Both fixatives yielded good tissue structure after autoclaving. In the sections fixed with formalin containing CaCl2, 20 of 22 antigens located in the nucleus, cytoplasm, membranes, and extracellular matrix greatly recovered their antigenicity after autoclaving; only two antigens exhibited stronger immunoreaction in acetone-fixed fresh frozen sections than these sections. Heating also retrieved the immunoreactivity of at least 14 antigens in the sections fixed with glutaraldehyde. We used the similar procedures to localize ligand-free estrogen receptor α (ERα) and glucocorticoid receptors (GR). Mouse uterine cells exhibited almost the same nuclear ERα immunostaining regardless of the hormonal status in glutaraldehyde-fixed fresh frozen sections and unliganded GR was localized mainly in the nucleus of mouse hepatocytes in fresh frozen sections fixed with 20% formalin containing 50 or 75 mM CaCl2 at 40C, after autoclaving. These results demonstrate that HIAR is useful for the immunohistochemistry of many antigens in aldehyde-fixed fresh frozen sections.

scholarly journals p-HYDROQUINONE AND p-BENZOQUINONE AS HISTOCHEMICAL REAGENTS I. A NEW TETRAZOLIUM METHOD FOR AMINO GROUPS, AND THE MECHANISM OF FORMAZAN STAINING OF PARAFFIN AND FRESH FROZEN SECTIONS

1967 ◽  
Vol 15 (7) ◽  
pp. 404-408 ◽  
Author(s):  
G. G. CARMICHAEL ◽  
STEPHANIE T. K. MANDER
Keyword(s):  
Electron Acceptor ◽  
Thermal Shrinkage ◽  
Frozen Sections ◽  
Amino Groups ◽  
Paraffin Sections ◽  
Tetrazolium Bromide ◽  
Acid Ph ◽  
Reduction System ◽  
Oxidation Reduction ◽  
Fresh Frozen

The staining of amino groups by formazan when dehydrated paraffin sections are incubated in a mixture of hydroquinone and 3-(4,5-dimethyl thiazolyl-2)-2 ,5-diphenyl-2H-tetrazolium bromide at an acid pH is reported. The mechanism of this reaction and of the cytoplasmic deposition of formazan in fresh frozen sections incubated under similar conditions is investigated. It is shown that the oxidation of hydroquinone to semiquinone is responsible for the reaction, the tetrazole acting as electron acceptor. The tissue amino groups, exposed by dehydration and thermal shrinkage, and the nitrogen groupings of phosphobipid behave as "catalysts." The relevant properties of the hydroquinone-benzoquinone oxidation-reduction system are described, and the reactions between benzoquinone and tissue constituents are reviewed.

scholarly journals A Molecular Mechanism of Formalin Fixation and Antigen Retrieval

2004 ◽  
Vol 121 (2) ◽  
pp. 190-199 ◽  
Author(s):  
Seshi R. Sompuram ◽  
Kodela Vani ◽  
Elizabeth Messana ◽  
Steven A. Bogen
Keyword(s):  
Molecular Mechanism ◽  
Formalin Fixation ◽  
Antigen Retrieval

Effect of bacterial endotoxemia on succinic dehydrogenase of liver and kidney of rabbit

1961 ◽  
Vol 201 (1) ◽  
pp. 16-18 ◽  
Author(s):  
J. Cascarano ◽  
A. D. Rubin ◽  
A. K. Neumann ◽  
B. W. Zweifach
Keyword(s):  
Succinic Dehydrogenase ◽  
Significant Inhibition ◽  
Frozen Sections ◽  
Experimental Animals ◽  
E Coli ◽  
Fresh Frozen ◽  
Tissue Homogenates ◽  
Liver And Kidney ◽  
Lethal Doses

The in vivo inhibition of liver and kidney succinic dehydrogenase by administration of lethal doses of bacterial endotoxin ( Escherichia coli and Salmonella typhosa) was investigated. Quantitative determinations conducted on tissue homogenates revealed significant inhibition of activity only in liver of rabbits injected with E. coli lipopolysaccharide. The histochemical distribution of succinic dehydrogenase in fresh frozen sections of kidney was the same in both control and experimental animals. However, the centrolobular areas of liver appeared considerably depressed in activity in both E. coli and S. typhosa endotoxin-treated animals. These data, along with those presented by other studies in the literature, suggest that the action of endotoxin appears to be restricted to certain cells.

Lipoproteins and Fibrinogen in Atherogenesis

1979 ◽  
Author(s):  
K.W. Walton
Keyword(s):  
Low Density Lipoproteins ◽  
Atherosclerotic Plaques ◽  
Low Density ◽  
Frozen Sections ◽  
Fibrin Monomer ◽  
Fresh Frozen ◽  
Arterial Intima

Previous work from this and other laboratories has shown that material antigenically related to the low-density lipoproteins (LDL) and to fibrinogen is demonstrable in atherosclerotic plaques. When arterial intima is extracted electrophoretically, ‘bound’ and ‘labile’ fractions of these antigens are in each case demonstrable. In the case of the “bound’ fraction of fibrinogen-related antigen (FRA) it has not been clear as to whether the material is in the form of native fibrinogen, as fibrin monomer, or as fibrin. To examine this problem fresh-frozen sections of arteries containing FRA have been examined by immunohistological techniques using antisera specific for fibrinogen, fibrinopeptides, plasmin and fibronectin. The results obtained with these antisera will be compared with one another and with results obtained with antisera to the antigens of LDL.

scholarly journals Granulomatous Pancreas: A Case Report of Pancreatic Sarcoid

2017 ◽  
Vol 2017 ◽  
pp. 1-3 ◽  
Author(s):  
Tatiana Bihun ◽  
Yanet Diaz ◽  
Seth Wenig
Keyword(s):  
Disease Process ◽  
Unknown Etiology ◽  
Frozen Sections ◽  
Gi Tract ◽  
Quadrant Pain ◽  
Pancreatic Involvement ◽  
Pancreatic Masses ◽  
Fresh Frozen ◽  
History Of ◽  
Head Of The Pancreas

Sarcoidosis is a chronic, systemic, noncaseating granulomatous disease process of unknown etiology. Sarcoidosis most commonly manifests in the lungs; however, gastrointestinal manifestations can occur. If in the GI tract, it is almost always found in the liver. Solitary pancreatic lesions are extremely rare, with less than 50 documented cases found in the literature. We present a case of a 61-year-old female, with a past medical history of sarcoidosis, who presented to the ER with unexpected weight loss, scleral icterus, right upper quadrant pain, and epigastric and back pain. US and MRI found a dilated common bile duct and mild dilation of the pancreatic duct, as well as a focal prominence in the head of the pancreas surrounded by areas of atrophy. A pancreaticoduodenectomy procedure was performed and fresh frozen sections were taken. The pathologist made a diagnosis of nonnecrotizing granulomatous pancreatitis. Pancreatic sarcoid is often asymptomatic and a benign finding on autopsy; however, clinicians should be mindful of pancreatic involvement when working up differential diagnosis for pancreatic masses.

scholarly journals Antigen Masking During Fixation and Embedding, Dissected

2016 ◽  
Vol 65 (1) ◽  
pp. 5-20 ◽  
Author(s):  
Carla Rossana Scalia ◽  
Giovanna Boi ◽  
Maddalena Maria Bolognesi ◽  
Lorella Riva ◽  
Marco Manzoni ◽  
...  
Keyword(s):  
Room Temperature ◽  
Detection System ◽  
Antigen Retrieval ◽  
Fixation Time ◽  
Water Removal ◽  
Immunohistochemical Detection ◽  
Frozen Sections ◽  
Ki 67 ◽  
Paraffin Embedding ◽  
Multiple Factors

Antigen masking in routinely processed tissue is a poorly understood process caused by multiple factors. We sought to dissect the effect on antigenicity of each step of processing by using frozen sections as proxies of the whole tissue. An equivalent extent of antigen masking occurs across variable fixation times at room temperature. Most antigens benefit from longer fixation times (>24 hr) for optimal detection after antigen retrieval (AR; for example, Ki-67, bcl-2, ER). The transfer to a graded alcohol series results in an enhanced staining effect, reproduced by treating the sections with detergents, possibly because of a better access of the polymeric immunohistochemical detection system to tissue structures. A second round of masking occurs upon entering the clearing agent, mostly at the paraffin embedding step. This may depend on the non-freezable water removal. AR fully reverses the masking due both to the fixation time and the paraffin embedding. AR itself destroys some epitopes which do not survive routine processing. Processed frozen sections are a tool to investigate fixation and processing requirements for antigens in routine specimens.

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