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Related Concept Videos

Immunocytochemistry and Immunohistochemistry01:22

Immunocytochemistry and Immunohistochemistry

10.6K
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.
These...
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Immunofluorescence Microscopy01:12

Immunofluorescence Microscopy

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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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Fixation and Sectioning01:03

Fixation and Sectioning

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Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens.
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...
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Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

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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.
4.0K
Simple Staining Technique01:24

Simple Staining Technique

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OverviewStaining techniques in microscopy enhance the visualization of microorganisms by increasing contrast and allowing the differentiation of cellular structures. Simple staining is one of the fundamental methods used to observe the basic morphological characteristics of microorganisms, including their size, shape, and arrangement. This method relies on the application of a single dye to stain the entire cell, producing a clear contrast between the cell and the background.FixationFixation is...
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Related Experiment Video

Updated: Jun 6, 2025

Histological-Based Stainings Using Free-Floating Tissue Sections
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Histological-Based Stainings Using Free-Floating Tissue Sections

Published on: August 25, 2020

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Brief guide to immunostaining.

Gyutae Park1, Sieun S Kim1, Jiwon Shim2

  • 1Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon 34141, South Korea.

Molecules and Cells
|November 21, 2024
PubMed
Summary
This summary is machine-generated.

This guide explains immunostaining, a key biological technique for visualizing antigen localization and levels in cells and tissues using antibodies. It covers principles, methods, types, and optimization strategies for researchers.

Keywords:
ImmunocytochemistryImmunofluorescenceImmunohistochemistry

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Immunohistochemistry on Paraffin Sections of Mouse Epidermis Using Fluorescent Antibodies
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Area of Science:

  • Biotechnology
  • Cell Biology
  • Immunology

Background:

  • Immunostaining is a fundamental technique in biological research.
  • It visualizes the location and quantity of specific molecules (antigens) within cells or tissues.
  • Antibodies are critical reagents for detecting target antigens.

Purpose of the Study:

  • To provide a concise guide to immunostaining techniques.
  • To explain the underlying principles and various methodologies.
  • To offer practical advice on common challenges and optimization.

Main Methods:

  • The manuscript outlines standard immunostaining protocols.
  • It details methods for antigen retrieval and antibody incubation.
  • Different detection systems and visualization strategies are discussed.

Main Results:

  • The guide covers direct and indirect immunostaining approaches.
  • It addresses fixation, permeabilization, and blocking steps.
  • Common pitfalls and troubleshooting tips are included.

Conclusions:

  • This resource aids researchers in understanding and performing immunostaining.
  • It enhances experimental design and data interpretation in biological studies.
  • Effective immunostaining is crucial for accurate molecular localization and quantification.