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Related Concept Videos

Immunofluorescence Microscopy01:12

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A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
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Immunocytochemistry and Immunohistochemistry01:22

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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

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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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Related Experiment Video

Updated: Aug 20, 2025

Indirect Immunofluorescence on Frozen Sections of Mouse Mammary Gland
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Immunohistochemistry and Immunofluorescence.

Haizal Mohd Hussaini1, Benedict Seo2, Alison M Rich2,3

  • 1School of Dentistry, University of Otago, Dunedin, New Zealand. haizal.mh@otago.ac.nz.

Methods in Molecular Biology (Clifton, N.J.)
|November 23, 2022
PubMed
Summary
This summary is machine-generated.

Immunohistochemistry (IHC) and immunofluorescence (IF) are vital protein detection methods in pathology. These techniques utilize specific antibody-antigen binding for visualizing biomarkers in tissues, aiding diagnosis and patient management.

Keywords:
Antigen-antibody complexDouble stainingImmunofluorescence (IF)Immunohistochemistry (IHC)Multiplex

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Optimization, Design and Avoiding Pitfalls in Manual Multiplex Fluorescent Immunohistochemistry
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Area of Science:

  • Biotechnology
  • Pathology
  • Immunology

Background:

  • Immunohistochemistry (IHC) is a cornerstone protein detection technique.
  • It relies on specific antibody-antigen interactions for visualization in tissues.
  • Enzymatic labels (peroxidase, alkaline phosphatase) or fluorescent signals (immunofluorescence, IF) are employed.

Purpose of the Study:

  • To review the principles of Immunohistochemistry (IHC).
  • To describe Immunofluorescence (IF) techniques.
  • To provide examples and outline multiplex/sequential staining protocols for IHC/IF.

Main Methods:

  • Utilizing specific antibodies to bind target antigens in tissue samples.
  • Employing enzymatic labels (e.g., peroxidase) or fluorescent signals for detection.
  • Application on formalin-fixed paraffin-embedded (FFPE) or fresh tissues; potential for multiplexing and sequential staining.

Main Results:

  • Demonstration of IHC and IF principles.
  • Examples of staining using common antibodies.
  • Description of double staining procedures for IHC/IF.

Conclusions:

  • IHC and IF are versatile techniques applicable to various tissue preparations.
  • These methods are crucial for diagnostic pathology, biomarker analysis, patient staging, and treatment decisions.
  • IF is particularly valuable for immune-mediated and vesiculobullous lesions in fresh biopsies.