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Related Concept Videos

Immunocytochemistry and Immunohistochemistry01:22

Immunocytochemistry and Immunohistochemistry

Immunocytochemistry (ICC) and immunohistochemistry (IHC) are techniques that use antibodies to check for specific proteins or antigens in a sample. The technique was first published by Albert Coons in 1941 to detect the presence of pneumococcal antigen in tissue sections from mice infected with Pneumococcus. Immunocytochemistry helps localization of proteins or antigens in individual cells like blood cells, stem cells, etc., while immunohistochemistry does the same for tissue samples.
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Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.

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Measurement of Four Uterine NK Cell Subtypes Using Multiplexed Fluorescent Immunohistochemical Staining in Women with Repeated Implantation Failure
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Multiple antigen immunostaining procedures.

Tibor Krenacs1, Laszlo Krenacs, Mark Raffeld

  • 11st Department of Pathology and Experimental Cancer Research, Semmelweis University, Budapest, Hungary.

Methods in Molecular Biology (Clifton, N.J.)
|December 17, 2009
PubMed
Summary
This summary is machine-generated.

Multiple antigen detection in tissues uses various immunohisto/cytochemical methods with chromogenic or fluorochrome labels. Protocols are discussed for simultaneous antibody application and avoiding cross-talk for precise cellular analysis.

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Area of Science:

  • Immunohistochemistry and Cytochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • Detecting multiple antigens within a single tissue section is crucial for comprehensive biological analysis.
  • Existing techniques often rely on chromogenic precipitates or fluorochrome labeling for visualization.

Purpose of the Study:

  • To discuss the principles of multiple antigen immunolabeling.
  • To provide practical protocols for performing multi-antigen detection in tissues.

Main Methods:

  • Utilizing combinations of immunohisto/cytochemical techniques, including immunogold silver staining, immunoperoxidase, and immunoalkaline phosphatase.
  • Employing various chromogens for distinct color precipitates and fluorochrome labels (e.g., AMCA, FITC, rhodamine, Cy5, Alexa series, quantum dots).
  • Strategies for simultaneous antibody application using directly labeled or non-cross-reacting antibodies.
  • Methods to prevent cross-talk when using cross-reacting antibodies, such as chromogen shielding, sequential inactivation (acidic elution, formaldehyde fixation, microwave heating), hapten labeling, and monovalent secondary antibodies.

Main Results:

  • Successful detection of multiple antigens in the same tissue section is achievable through diverse immunolabeling strategies.
  • Specific protocols enable the differentiation of antigens located in separate cellular compartments or cells.

Conclusions:

  • Multiple antigen immunolabeling is a versatile technique for detailed cellular and tissue analysis.
  • The discussed methods and protocols offer solutions for complex multi-antigen detection challenges, enhancing diagnostic and research capabilities.