Immunolabeling
Immunolabeling - Related Videos
02:19
Immunolabeling of Neuronal Membrane Proteins in a Freeze-fractured Specimen of Mouse Brain Tissue
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2025
Source: Schönherr, S., et. al. Combined Optogenetic and Freeze-fracture Replica Immunolabeling to Examine Input-specific Arrangement of Glutamate Receptors in the Mouse Amygdala. J. Vis. Exp. (2016)This video demonstrates the immunolabelling of freeze-fractured replicas of mouse brain tissue for electron microscopy. The fractured and coated replicas are digested to expose receptors. Primary and gold-tagged secondary antibodies are added to label specific neuronal membrane receptors. The replica...
09:49
Combined Optogenetic and Freeze-fracture Replica Immunolabeling to Examine Input-specific Arrangement of Glutamate Receptors in the Mouse Amygdala
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Cited by 6 •
2016
This article illustrates how the expression of neurotransmitter receptors can be quantified and the pattern analyzed at synapses with identified pre and postsynaptic elements using a combination of viral transduction of optogenetic tools and the freeze-fracture replica immunolabeling technique.
06:18
Optical Clearing and Imaging of Immunolabeled Kidney Tissue
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Cited by 7 •
2019
The combination of antibody labeling, optical clearing, and advanced light microscopy allows three-dimensional analysis of complete structures or organs. Described here is a simple method to combine immunolabeling of thick kidney slices, optical clearing with ethyl cinnamate, and confocal imaging that enables visualization and quantification of three-dimensional kidney structures.
06:37
Immunolabelling Myofiber Degeneration in Muscle Biopsies
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Cited by 16 •
2019
Described here is a protocol for direct immunolabelling of necrotic myofibers in muscle cryosections. Necrotic cells are permeable to serum proteins, including immunoglobulin G (IgG). Revealing the uptake of IgG by myofibers allows the identification and quantification of myofibers that undergo necrosis regardless of muscle condition.
05:04
An Immunolabeling Technique to Visualize the Subcellular Localization of Proteins
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2025
This video demonstrates a technique for the immunolabeling of sigma-1 receptors in a murine heart tissue section using quantum dots. The heart tissue is fixed, osmicated, stained, dehydrated, embedded in resin, and then sectioned. The sections are then treated with antibodies and labeled with quantum dots to demarcate the subcellular localization of Sigma-1 receptors using electron microscopy.
FAQs
The primary antibody provides target recognition by binding the selected protein or antigen, whereas the secondary antibody supplies the detectable label when an indirect strategy is used. This separation allows the same target-binding step to be paired with a fluorescent or enzymatic signal. Directly conjugated detection reagents can instead provide the signal without a labeled secondary antibody.
Fixation and permeabilization support different requirements of the assay. Fixation helps preserve the organization of cells and tissues, maintaining the structural context in which neuronal markers are interpreted. Permeabilization helps antibodies reach targets within the sample. Together, these preparation steps allow molecular detection while retaining tissue architecture for microscopy.
Fluorescent detection produces signals that can be visualized by fluorescence microscopy, while enzymatic detection uses a labeled reagent that generates an observable enzymatic signal. The choice affects how the labeled target is examined in the sample. In neuroscience, either approach can support analysis of protein distribution and the location of neuronal markers within tissue.
A typical workflow begins by preparing the sample with fixation and permeabilization, then exposing it to a primary antibody directed against the selected neuronal marker or other antigen. A labeled secondary antibody or directly conjugated detection reagent is subsequently used to generate the signal. Microscopy then reveals the marker’s distribution in its cellular or tissue context.
Multiplex immunolabeling is useful when researchers need to examine several cellular components in the same complex neural sample. Simultaneous labeling can show how different markers are distributed relative to one another, supporting the study of cellular relationships within neural circuits. This approach can help connect individual marker patterns with broader circuit organization.
In neuroscience, immunolabeling can map neuronal markers, distinguish cell types, and reveal where particular proteins occur within cells or tissues. Comparing labeling patterns across samples can also support studies of development, disease, or injury. These outcomes link molecular information to neural structure and provide microscopy-based evidence about changes in biological samples.