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Related Concept Videos

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.
Antigen Processing Pathways01:31

Antigen Processing Pathways

MHC molecules are key players in the immune response, enabling T cells to recognize and respond to specific antigens. They are present on the surface of all nucleated cells in the body and are instrumental in presenting antigens to T cells and activating them. T cells recognize the MHC-antigen complex and initiate an immune response. MHC class I and MHC class II are two main types of MHC molecules, each associated with a distinct antigen processing pathway.
MHC Class I: Presenting Endogenous...
Antigen Presenting Cells01:22

Antigen Presenting Cells

The immune system is a complex network of cells and molecules that protects the body from foreign invaders. T cells, a type of white blood cell, play a crucial role in this process. They recognize and attack foreign substances, such as pathogens, that enter the body.
T cells require the help of antigen-presenting cells (APCs), which process foreign antigens into smaller fragments that can be recognized by T cells. These APCs are highly specialized cells that efficiently internalize antigens...
Antigens Involved in Adaptive Immunity01:26

Antigens Involved in Adaptive Immunity

An antigen is any substance the immune system identifies as foreign and potentially harmful to the body, prompting an immune response. Antigens have two functional properties: immunogenicity and reactivity. Immunogenicity is the ability of an antigen to stimulate a specific immune response. At the same time, reactivity describes the antigen's ability to react with the cells and antibodies produced in response to it.
Complete Antigens
Complete antigens possess both immunogenicity and reactivity.

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Isolation And Dendritic Cell-Uptake of Small Extracellular Vesicles from Echinococcus granulosus
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Immunoelectron microscopy of antigen processing in dendritic cells.

M J Kleijmeer1, G Posthuma, H J Geuze

  • 1Department of Cell Biology and Institute of Biomembranes, Faculty of Medicine, Utrecht University, The Netherlands.

Methods in Molecular Medicine
|March 5, 2011
PubMed
Summary

High-resolution immunoelectron microscopy (IEM) using immunogold labeling on cryosections is optimal for studying intracellular compartments. This method preserves ultrastructure and antigens for detailed cell biology research.

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Area of Science:

  • Cell Biology
  • Microscopy Techniques

Background:

  • Understanding intracellular compartments and their protein interactions is crucial in cell biology.
  • High-resolution immunoelectron microscopy (IEM) is a key technique for this investigation.

Purpose of the Study:

  • To detail the methodology of immunoelectron microscopy (IEM) for studying intracellular compartments.
  • To highlight the advantages of IEM, particularly immunogold labeling on cryosections.

Main Methods:

  • Utilizing immunogold labeling on cryosections for IEM.
  • Preparation of ultrathin cryosections (50-100 nm) from tissue blocks or cell pellets.
  • Antibody labeling visualized with protein A-gold particles (PAG).

Main Results:

  • IEM with immunogold labeling on cryosections provides optimal ultrastructure.
  • This technique ensures good preservation of protein and/or lipid antigens.
  • It offers better accessibility of immunodeterminants to antibodies compared to other methods.

Conclusions:

  • Immunoelectron microscopy (IEM) is the optimum technique for detailed cell biology studies of intracellular compartments.
  • Cryosectioning combined with immunogold labeling offers superior antigen and ultrastructure preservation.
  • The described methods provide a robust approach for visualizing protein localization and interactions within cells.