Postembedding Immunogold
Postembedding Immunogold - Related Videos
09:24
High-Resolution Quantitative Immunogold Analysis of Membrane Receptors at Retinal Ribbon Synapses
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Cited by 7 •
2016
The postembedding immunogold method is one of the most effective ways to provide high-resolution analyses of the subcellular localization of specific molecules. Here we describe a protocol to quantitatively analyze glutamate receptors at retinal ribbon synapses.
01:20
Immunogold Electron Microscopy
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2025
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.
13:50
Post-embedding Immunogold Labeling of Synaptic Proteins in Hippocampal Slice Cultures
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Cited by 13 •
2013
The localization and distribution of proteins provide important information for understanding their cellular functions. The superior spatial resolution of electron microscopy (EM) can be used to determine the subcellular localization of a given antigen following immunohistochemistry. For tissues of the central nervous system (CNS), preserving structural integrity while maintaining antigenicity has been especially difficult in EM studies. Here, we adopt a procedure that has been used to preserve...
09:49
Nanogold Labeling of the Yeast Endosomal System for Ultrastructural Analyses
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Cited by 1 •
2014
Yeast, Saccharomyces cerevisiae, has been a key model organism to identify and study genes regulating the biogenesis and functions of the endosomal system. Here we present a detailed protocol for the specific labeling of the endosomal compartments for ultrastructural studies.
07:30
Measuring Synaptic Vesicle Endocytosis in Cultured Hippocampal Neurons
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Cited by 10 •
2017
Synaptic vesicle endocytosis is detected by light microscopy of pHluorin fused with synaptic vesicle protein and by electron microscopy of vesicle uptake.
FAQs
Antibodies bind target molecules that are exposed on the surface of the resin-embedded, ultrathin section. Consequently, the observed signal reflects accessible antigen at that cut surface rather than every molecule present throughout the tissue. This surface-dependent labeling is important when interpreting whether a neurotransmitter, receptor, or synaptic protein occupies a particular cellular or synaptic location.
A secondary antibody carries colloidal gold particles that are electron-dense, so they appear as visible markers in transmission electron microscopy. The particles convert an antibody-antigen interaction into a localized ultrastructural signal. Their position can therefore be examined alongside neuronal membranes, synaptic regions, and subcellular compartments, linking molecular labeling with nanometer-scale structural context.
Immunogold labeling places electron-dense markers relative to the fine structure of a synapse. Comparing particle locations with presynaptic and postsynaptic regions allows researchers to determine whether a neurotransmitter, receptor, ion channel, or synaptic protein is concentrated on one side or distributed across both. This spatial information helps relate molecular organization to synaptic transmission and plasticity.
The workflow proceeds from tissue embedding in resin to preparation of ultrathin sections, followed by antibody binding to exposed target molecules on the section surface. A gold-particle-conjugated secondary antibody then marks the bound primary antibody, and transmission electron microscopy visualizes the electron-dense particles. The resulting images combine antigen localization with the ultrastructure of the section.
In neuroscience, the approach can localize neurotransmitters, receptors, ion channels, and other synaptic proteins within neurons and their subcellular compartments. Examining these targets at the same ultrastructural scale helps researchers map where molecular components occur in relation to synapses. Such maps support studies of synaptic organization and the molecular basis of neuronal communication.
The localization patterns can be used to investigate how molecular components are arranged within synapses and other neuronal compartments. Researchers can relate the distribution of labeled targets to synaptic transmission, plasticity, and neurological disease. Because the method provides structural context at nanometer scale, it connects changes in protein or neurotransmitter location with specific cellular sites.