Localization of transplantation antigens in tissue sections: Effects of various fixatives and use of tissue preparations other than frozen sections

1972 ◽  
Vol 28 (3) ◽  
pp. 342-343 ◽  
Author(s):  
A. G. Gervais
Keyword(s):  
Frozen Sections ◽  
Tissue Sections ◽  
Transplantation Antigens

scholarly journals Magnetically Promoted Rapid Immunofluorescence Staining for Frozen Tissue Sections

2019 ◽  
Vol 67 (8) ◽  
pp. 575-587 ◽  
Author(s):  
Tatsuya Onishi ◽  
Sachiko Matsuda ◽  
Yuki Nakamura ◽  
Junko Kuramoto ◽  
Akinori Tsuruma ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Growth Factor Receptor ◽  
Metastatic Breast ◽  
Detection Methods ◽  
Frozen Sections ◽  
Tissue Sections ◽  
Tissue Arrays ◽  
Abnormal Cells ◽  
Epidermal Growth ◽  
Hematoxylin And Eosin

Current immunohistochemistry methods for diagnosing abnormal cells, such as cancer cells, require multiple steps and can be relatively slow compared with intraoperative frozen hematoxylin and eosin staining, and are therefore rarely used for intraoperative examination. Thus, there is a need for novel rapid detection methods. We previously demonstrated that functionalized fluorescent ferrite beads (FF beads) magnetically promoted rapid immunoreactions. The aim of this study was to improve the magnetically promoted rapid immunoreaction method using antibody-coated FF beads and a magnet subjected to a magnetic field. Using frozen sections of xenograft samples of A431 human epidermoid cancer cells that express high levels of epidermal growth factor receptor (EGFR) and anti-EGFR antibody-coated FF beads, we reduced the magnetically promoted immunohistochemistry procedure to a 1-min reaction and 1-min wash. We also determined the optimum magnetic force for the antibody reaction (from 7.79 × 10−15 N to 3.35 × 10−15 N) and washing (4.78 × 10−16 N), which are important steps in this technique. Furthermore, we stained paraffin-embedded tissue arrays and frozen sections of metastatic breast cancer lymph nodes with anti-pan-cytokeratin antibody-coated FF beads to validate the utility of this system in clinical specimens. Under optimal conditions, this ultra-rapid immunostaining method may provide an ancillary method for pathological diagnosis during surgery. (J Histochem Cytochem 58:XXX–XXX, 2010)

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.

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 A postcoupling method for the demonstration of N-acetyl-beta-D-glucosaminidase in unfixed frozen tissue sections.

1975 ◽  
Vol 23 (6) ◽  
pp. 424-430 ◽  
Author(s):  
A D Shannon
Keyword(s):  
Enzyme Activity ◽  
Incubation Medium ◽  
Present Method ◽  
Diazonium Salts ◽  
Staining Reaction ◽  
Tissue Fixation ◽  
Frozen Sections ◽  
Inhibitory Effects ◽  
Tissue Sections ◽  
The Comparative Study

The localization and activity of the lysosomal enzyme, N-acetyl-beta-glucosaminidase, has been studied in unfixed frozen sections of tissues from normal and hemorrhaged rats with the use of a modified postcoupling technique. Discrete localization of the end product of the reaction was achieved in this method by the incorporation of 20% (w/v) polyvinyl alcohol (molecular weight 14,000) in the incubation medium. Advantages of the present method include the ability to overcome the inhibitory effects on enzyme activity of both tissue fixation and the presence of diazonium salts in the incubation medium. The staining reaction obtained with this technique demonstrates the enzyme activity at specific cytoplasmic sites within cells and has a wider applicability to the comparative study of N-acetyl-beta-glucosaminidase activity in normal and injured tissues.

scholarly journals Light microscopic visualization of colloidal gold on resin-embedded tissue.

1983 ◽  
Vol 31 (12) ◽  
pp. 1394-1398 ◽  
Author(s):  
G Danscher ◽  
J O Nörgaard
Keyword(s):  
Colloidal Gold ◽  
Microscopic Level ◽  
Electron Microscopic Level ◽  
Metallic Silver ◽  
Frozen Sections ◽  
Electron Microscopic ◽  
Tissue Sections ◽  
Amplification Method ◽  
Glass Slides ◽  
Present Technique

RNase labeled with colloidal gold was used as a model for the present technique evolved for the light microscopic localization of gold-labeled substances in semithin resin-embedded sections. Tissue sections placed on glass slides were treated with the gold-enzyme complex and subsequently exposed to a photographic developed containing silver lactate. During the development gold particles are encapsulated in growing shells of metallic silver and gradually made visible in the light microscope. The amplification method can be applied to paraffin-embedded and frozen sections as well. This technique may prove useful as a supplement to studies utilizing colloidal gold or silver as markers normally used at the electron microscopic level.

scholarly journals Immunofluorescent staining of histaminase (diamine oxidase) in human placenta.

1978 ◽  
Vol 26 (11) ◽  
pp. 1021-1025 ◽  
Author(s):  
C W Lin ◽  
C M Chapman ◽  
R A DeLellis ◽  
S Kirley
Keyword(s):  
Intercellular Space ◽  
Diamine Oxidase ◽  
Intracellular Localization ◽  
Immunofluorescent Staining ◽  
Frozen Sections ◽  
Chorionic Villi ◽  
Tissue Sections ◽  
Decidual Cells ◽  
Tissue Space

An immunofluorescent procedure for the localization of histaminase in human tissue sections has been developed by using a specific antiserum against human placental histaminase. For localization of this enzyme in placental sections, fixation in equal volumes mixture of absolute ethanol and acetone provided the optimum visualization of this enzyme in both frozen sections and paraffin-embedded sections. The immunofluorescent staining of this enzyme in placenta is found to be localized in areas within the maternal decidua, both within the cytoplasm of the decidual cells and in tissue space between the cells. The chorionic villi are completely void of the immunofluorescent stain. Variations in patterns of histaminase localization have been found between term and premature placentas, with the former showing a predominantly intercellular localization and the latter a predominantly intracellular localization. The intercellular localization of this enzyme in the decidua may represent a nonspecific diffusion of the enzyme associated with delivery of the placenta or may reflex a specific functional role of the enzyme in the intercellular space during pregnancy.

scholarly journals Quenching autofluorescence in tissue immunofluorescence

2017 ◽  
Vol 2 ◽  
pp. 79 ◽  
Author(s):  
Jian Yang ◽  
Fengtang Yang ◽  
Lia S. Campos ◽  
William Mansfield ◽  
Helen Skelton ◽  
...  
Keyword(s):  
Pluripotent Stem Cells ◽  
Poor Quality ◽  
Frozen Sections ◽  
Combined Treatments ◽  
Tissue Sections ◽  
Sudan Black B ◽  
Specific Proteins ◽  
Teratoma Formation ◽  
Texas Red ◽  
Embryonic Organ

Background: Immunofluorescence (IF) is one of the most important techniques where fluorochromes conjugated to antibodies are used to detect specific proteins or antigens. In tissue sections, autofluorescence (AF) can lead to poor quality images that impair assessment. The placenta is a pivotal extra-embryonic organ in embryo development, where trophoblasts make up a large proportion of the cells. Teratoma formation is one of the critical assays for pluripotent stem cells. Methods: We tested whether ultraviolet (UV), ammonia (NH3), copper (II) sulfate (CuSO4), Trypan Blue (TB), Sudan Black B (SB), TrueBlack™ Lipofusin Autofluorescence Quencher (TLAQ) and combinations of these treatments could reduce AF in paraffin and frozen sections of placenta and teratoma in FITC, Texas Red and Cy5.5 channels. Results: We found that UV, NH3, TB and CuSO4 quenched AF to some extent in different tissue and filters, but increased AF in Texas Red or Cy5.5 channels in some cases. SB and TLQA exhibited the most consistent effects on decreasing AF, though TLQA reduced the overall IF signal in placenta sections. Not all combined treatments further reduced AF in both placenta and teratoma sections. Conclusions: SB and TLAQ can effectively quench AF in placenta and teratoma IF.

scholarly journals Bringing SEM and MSI Closer Than Ever Before: Visualizing Aspergillus and Pseudomonas Infection in the Rat Lungs

2020 ◽  
Vol 6 (4) ◽  
pp. 257
Author(s):  
Tereza Juříková ◽  
Dominika Luptáková ◽  
Olga Kofroňová ◽  
Anton Škríba ◽  
Jiří Novák ◽  
...  
Keyword(s):  
Mass Spectrometry Imaging ◽  
Ionization Mass ◽  
Bacterial Cells ◽  
Laser Desorption Ionization ◽  
Rat Lung ◽  
Frozen Sections ◽  
Molecular Biomarker ◽  
Tissue Sections ◽  
Pseudomonas Infection ◽  
Ethanol Series

A procedure for processing frozen rat lung tissue sections for scanning electron microscopy (SEM) from deeply frozen samples initially collected and stored for matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) was developed. The procedure employed slow thawing of the frozen sections while floating on the surface and melting in a fixative solution. After the float-washing step, the sections were dehydrated in a graded ethanol series and dried in a critical point dryer. The SEM generated images with well-preserved structures, allowing for monitoring of bacterial cells and fungal hyphae in the infected tissue. Importantly, the consecutive nonfixed frozen sections were fully compatible with MALDI-MSI, providing molecular biomarker maps of Pseudomonas aeruginosa. The protocol enables bimodal image fusion in the in-house software CycloBranch, as demonstrated by SEM and MALDI-MSI.

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.

Modifications to the Gomori Acid Phosphatase Technique for Controlled-Temperature Frozen Sections

1963 ◽  
Vol s3-104 (66) ◽  
pp. 193-196
Author(s):  
LUCILLE BITENSKY
Keyword(s):  
Aqueous Solution ◽  
Acetic Acid ◽  
Acid Phosphatase ◽  
Phosphatase Activity ◽  
Acid Medium ◽  
Hydrogen Sulphide ◽  
Acid Phosphatase Activity ◽  
Distilled Water ◽  
Frozen Sections ◽  
Tissue Sections

The preservation of inert lysosomes in tissue sections depends on the use of the controlled-temperature freezing-sectioning technique. The Gomori procedure for acid phosphatase produces considerable disintegration of these unfixed sections. This disintegration is not due to incubation in the acid medium nor to the rinsing either in dilute acetic acid or in distilled water, but to the treatment with the solution of ammonium sulphide. It is suggested that for this solution there should be substituted a saturated aqueous solution of hydrogen sulphide gas, which does not cause such cellular deformation. Another improvement involves the deletion of the rinse in acetic acid, because this might render soluble some of the specific precipitate in sections showing minimal acid phosphatase activity.

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