scholarly journals Mixed glycosidase pretreatment reduces nonspecific binding of antibodies to frozen tissue sections.

1985 ◽  
Vol 33 (7) ◽  
pp. 695-698 ◽  
Author(s):  
L P Andrews ◽  
R K Clark ◽  
I Damjanov
Keyword(s):  
Monoclonal Antibody ◽  
Renal Tissue ◽  
Frozen Sections ◽  
Tissue Sections ◽  
Mouse Tissues ◽  
Background Staining ◽  
Frozen Tissue ◽  
Nonspecific Staining ◽  
Immunohistochemical Studies ◽  
Monoclonal Antibody Binding

Indirect immunohistochemical studies of frozen mouse tissues with mouse monoclonal antibodies yield, in general, suboptimal results primarily because of indiscriminate binding of secondary antibody to all mouse immunoglobulins, i.e., to the monoclonal reagent and to endogenous immunoglobulin nonspecifically trapped in the tissue. To reduce this nonspecific staining, frozen sections of mouse kidney were treated enzymatically. Optimal results were obtained following a 2 hr treatment with 20 mg/ml of mixed glycosidases (MG). This treatment reduced the nonspecific background staining of the interstitial spaces and blood vessels, but did not affect the reactivity of structurally bound immunoglobulin G (IgG) in the glomeruli or alter the reactivity of mouse renal tissue to the monoclonal antibody that recognizes an oligosaccharide antigenic determinant (SSEA-1). Eluates from enzyme-treated frozen tissue sections contained normally immunoreactive IgG in the form of dimers. These data indicate that MG treatment of frozen sections could be safely used to reduce the content of nonstructurally bound immunoglobulins in frozen tissues and thus improve the visualization of specific monoclonal antibody binding.

scholarly journals Monoclonal antibodies to laminin reveal the heterogeneity of basement membranes in the developing and adult mouse tissues.

1984 ◽  
Vol 98 (3) ◽  
pp. 971-979 ◽  
Author(s):  
Y J Wan ◽  
T C Wu ◽  
A E Chung ◽  
I Damjanov
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Adult Mouse ◽  
Basement Membranes ◽  
Newborn Mouse ◽  
Preimplantation Stage ◽  
Mouse Tissues ◽  
Reichert's Membrane ◽  
Anatomic Locations ◽  
Immunohistochemical Studies

Two monoclonal antibodies raised against laminin isolated from a mouse parietal yolk sac cell line were used for immunohistochemical studies of basement membranes of the mouse embryo and various fetal and adult tissues. No immunoreactivity with either of the two monoclonal antibodies could be detected in the preimplantation-stage embryos, although it has been shown that these embryos contain extracellular laminin reactive with the conventional polyclonal antilaminin antibodies. Reichert's membrane in early postimplantation stages of development reacted with the monoclonal antibody LAM-I but not with the antibody LAM-II. However, from day 8 of pregnancy onward the Reichert's membrane reacted with both antibodies. Basement membranes of the embryo proper were unreactive with both monoclonal antibodies until day 12 of pregnancy. By day 14 some basement membranes of the fetal tissues became reactive with one or both monoclonal antibodies, whereas others remained still unreactive. In the 17-d fetus and the newborn mouse most of the basement membranes reacted with both monoclonal antibodies, whereas others still reacted with only one. Similar heterogeneity in the immunoreactivity of basement membranes of various tissues was noted in the adult mouse as well. These results indicate that the immunoreactivity of laminin in the extracellular matrix changes during development and that the basement membranes in various anatomic locations display heterogeneity even in the adult mouse.

The use of frozen tissue for diagnostic electron microscopy

1984 ◽  
Vol 42 ◽  
pp. 58-59
Author(s):  
R. Van de Velde ◽  
K. Chien ◽  
R. Heusser
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Sample Size ◽  
Needle Biopsy ◽  
Renal Tissue ◽  
Frozen Sections ◽  
Time Of Surgery ◽  
Frozen Tissue ◽  
Work Up

Specimens received for evaluation by a diagnostic electron microscopy laboratory do not always meet the accepted criteria for optimal preservation. Several reasons can be cited including delayed fixation, improper fixative or sample size. When a biopsy containing the diagnostic lesion is frozen at the time of surgery for an immediate diagnosis based on a cryosection or when one has a small biopsy and is not able to set aside sufficient tissue for all study parameters electron microscopy is often deleted. In our laboratory when a needle biopsy of kidney is insufficient for a complete microscopic work-up a decision may be made to freeze the tissue to provide rapid immunohistochemical data. After sufficient frozen sections are cut, the frozen tissue can then be placed in fixative for light microscopy. Because ultrastructural data sometimes would be necessary for a complete diagnosis, we have used frozen sections of renal tissue for EM studies as well.

Mucin Stain on Frozen Sections

1999 ◽  
Vol 123 (5) ◽  
pp. 378-380
Author(s):  
Suchetha Soans ◽  
Lorenzo M. Galindo ◽  
Fernando U. Garcia
Keyword(s):  
Colonic Mucosa ◽  
Frozen Section ◽  
Staining Technique ◽  
Turnaround Time ◽  
Staining Procedure ◽  
Normal Colonic Mucosa ◽  
Frozen Sections ◽  
Tissue Sections ◽  
Paget Disease ◽  
Frozen Tissue

Abstract Context.—The importance of frozen-section diagnoses in the practice of pathology cannot be overemphasized. In some cases, the use of a mucin stain can greatly aid in the diagnosis. Since few methods for mucin staining have been described that can be used in the frozen-section setting, we developed one such staining procedure for mucin. Objective.—To develop a rapid mucicarmine staining technique to be used on frozen sections that does not significantly delay overall turnaround time. Design.—A standard mucicarmine staining technique was modified by using a concentrated mucicarmine stain and a microwave oven, to reduce the total staining time to 3 minutes or less. Frozen tissue from normal colonic mucosa was used as a control, and skin from extramammary Paget disease for evaluation of margins was used as a case. Results.—The rapid mucicarmine stain successfully demonstrated the presence of mucin on frozen-tissue sections. Mucin stained deep rose, and the connective tissue stained green. Conclusion.—This rapid and simple mucin staining technique can be used on frozen sections with no significant effect on the overall turnaround time, thereby aiding rapid diagnosis on frozen sections.

Immunohistochemical demonstration of estrophilin in mouse tissues using a biotinylated monoclonal antibody.

1988 ◽  
Vol 36 (12) ◽  
pp. 1527-1531 ◽  
Author(s):  
J Tse ◽  
S Goldfarb
Keyword(s):  
Monoclonal Antibody ◽  
No Value ◽  
Mouse Liver ◽  
Immunohistochemical Method ◽  
Tissue Sections ◽  
Immunohistochemical Demonstration ◽  
Mouse Tissues ◽  
Microscopic Demonstration ◽  
Subsequent Exposure

We have developed a direct avidin-biotin-peroxidase complex (ABC) immunohistochemical method for localization of estrophilin in mouse tissues. The method has been found especially useful for microscopic demonstration of the receptor in mouse liver, since the indirect alternative, autoradiography after injection of radiolabeled estrogens, is of no value in this organ. The ABC technique employs a biotinylated monoclonal antibody to human estrophilin (Abbot H222) which was previously shown to crossreact with the murine receptor. Cryostat-cut tissue sections which were briefly fixed were incubated with the modified antibody, and the estrophilin was revealed by subsequent exposure to ABC followed by H2O2/diaminobenzidine.

scholarly journals Characterization of a Murine Monoclonal Antibody toCryptococcus neoformans Polysaccharide That Is a Candidate for Human Therapeutic Studies

1998 ◽  
Vol 42 (6) ◽  
pp. 1437-1446 ◽  
Author(s):  
Arturo Casadevall ◽  
Wendy Cleare ◽  
Marta Feldmesser ◽  
Aharona Glatman-Freedman ◽  
David L. Goldman ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Biological Properties ◽  
Murine Monoclonal Antibody ◽  
Preclinical Development ◽  
Antigen Binding Site ◽  
Therapeutic Trials ◽  
Mouse Tissues ◽  
Normal Human ◽  
Rapid Clearance ◽  
Immunohistochemical Studies

ABSTRACT The murine monoclonal antibody (MAb) 18B7 [immunoglobulin G1(κ)] is in preclinical development for treatment ofCryptococcus neoformans infections. In anticipation of its use in humans, we defined the serological and biological properties of MAb 18B7 in detail. Structural comparison to the related protective MAb 2H1 revealed conservation of the antigen binding site despite several amino acid differences. MAb 18B7 was shown by immunofluorescence and agglutination studies to bind to all four serotypes of C. neoformans, opsonize C. neoformans serotypes A and D, enhance human and mouse effector cell antifungal activity, and activate the complement pathway leading to deposition of complement component 3 (C3) on the cryptococcal capsule. Administration of MAb 18B7 to mice led to rapid clearance of serum cryptococcal antigen and deposition in the liver and spleen. Immunohistochemical studies revealed that MAb 18B7 bound to capsular glucuronoxylomannan in infected mouse tissues. No reactivity of MAb 18B7 with normal human, rat, or mouse tissues was detected. The results show that both the variable and constant regions of MAb 18B7 are biologically functional and support the use of this MAb in human therapeutic trials.

Study of vitrified, unstained frozen tissue sections by cryoimmunoelectron microscopy

1991 ◽  
Vol 100 (1) ◽  
pp. 227-236
Author(s):  
I. Sabanay ◽  
T. Arad ◽  
S. Weiner ◽  
B. Geiger
Keyword(s):  
Heavy Metal ◽  
High Resolution ◽  
High Contrast ◽  
Frozen Sections ◽  
Microscope Examination ◽  
Tissue Sections ◽  
Novel Approach ◽  
Cytoplasmic Filaments ◽  
Frozen Tissue ◽  
Ultrathin Frozen Sections

We describe the development and application of a novel approach to high-resolution ultrastructural analysis of cells and tissues. It is based on the preparation of ultrathin frozen sections of fixed tissues, rinsing of the sections, followed by their embedding on the grid in a layer of vitrified ice, and direct observation with a cryoelectron microscope. Examination of smooth muscle, kidney and heart tissues showed that although no heavy metal staining was used, high-contrast images are obtained. Fine details of cytoplasmic filaments and organelles, membranes and membrane-associated structures, as well as connective-tissue elements are all visible. The new method is suitable for immunolabeling, including high resolution localization of specific molecules within the cytoplasm.

scholarly journals The labeled antigen method of immunoenzymatic staining.

1979 ◽  
Vol 27 (4) ◽  
pp. 832-840 ◽  
Author(s):  
D Y Mason ◽  
R E Sammons
Keyword(s):  
Alkaline Phosphatase ◽  
Horseradish Peroxidase ◽  
Tissue Section ◽  
Specific Antibody ◽  
High Efficiency ◽  
Two Stage ◽  
Tissue Sections ◽  
Autoimmune Antibodies ◽  
Background Staining ◽  
Nonspecific Staining

A two stage immunohistological technique (the "labeled antigen" procedure) has been assessed for the detection of a variety of human and animal cytoplasmic constituents in tissue sections. In this method specific antiserum is followed by antigen complexed to horseradish peroxidase or to alkaline phosphatase. The primary antibody acts bivalently, linking the labeled antigen to antigen in the tissue section. The major advantage of this technique is that nonantigen specific antibody in the primary antiserum cannot cause nonspecific staining since it has no affinity for the antigen:enzyme complex. Consequently the specificity of the reaction is assured, background staining is minimized and the total staining time (from wax section to mounted slide) can be reduced to as little as 30 min. Further advantages include the possibility of labeling Ig allotypes and the high efficiency of enzyme utilization. Covalent human IgG:horseradish peroxidase complexes can also be used in a triple sandwich in conjunction with human anti-viral or autoimmune antibodies.

IgE and monoclonal antibody binding by the mite allergen Der p 7

1996 ◽  
Vol 26 (3) ◽  
pp. 308-315 ◽  
Author(s):  
H.-D. SHEN ◽  
K. Y. CHUA ◽  
W. L. LIN ◽  
H. L. CHEN ◽  
K.-H. HSIEH ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Mite Allergen ◽  
Antibody Binding ◽  
Monoclonal Antibody Binding

scholarly journals Rapid brain structure and tumour margin detection on whole frozen tissue sections by fast multiphotometric mid-infrared scanning

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Tim Kümmel ◽  
Björn van Marwick ◽  
Miriam Rittel ◽  
Carina Ramallo Guevara ◽  
Felix Wühler ◽  
...  
Keyword(s):  
Imaging System ◽  
Frozen Section ◽  
Computational Effort ◽  
Primary Hepatocellular Carcinoma ◽  
Measuring System ◽  
Sufficient Information ◽  
Tissue Samples ◽  
Mid Infrared ◽  
Tissue Sections ◽  
Frozen Tissue

AbstractFrozen section analysis is a frequently used method for examination of tissue samples, especially for tumour detection. In the majority of cases, the aim is to identify characteristic tissue morphologies or tumour margins. Depending on the type of tissue, a high number of misdiagnoses are associated with this process. In this work, a fast spectroscopic measurement device and workflow was developed that significantly improves the speed of whole frozen tissue section analyses and provides sufficient information to visualize tissue structures and tumour margins, dependent on their lipid and protein molecular vibrations. That optical and non-destructive method is based on selected wavenumbers in the mid-infrared (MIR) range. We present a measuring system that substantially outperforms a commercially available Fourier Transform Infrared (FT-IR) Imaging system, since it enables acquisition of reduced spectral information at a scan field of 1 cm2 in 3 s, with a spatial resolution of 20 µm. This allows fast visualization of segmented structure areas with little computational effort. For the first time, this multiphotometric MIR system is applied to biomedical tissue sections. We are referencing our novel MIR scanner on cryopreserved murine sagittal and coronal brain sections, especially focusing on the hippocampus, and show its usability for rapid identification of primary hepatocellular carcinoma (HCC) in mouse liver.

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