Post-embedding Technique
Post-embedding Technique - Related Videos
13:50
Post-embedding Immunogold Labeling of Synaptic Proteins in Hippocampal Slice Cultures
0 Views •
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...
08:47
Epon Post Embedding Correlative Light and Electron Microscopy
0 Views •
Cited by 1 •
2024
We present a detailed protocol for Epon post-embedding correlative light and electron microscopy using a fluorescent protein called mScarlet. This method can maintain the fluorescence and the ultrastructure simultaneously. This technique is amenable to a wide variety of biological applications.
09:17
Minimally Invasive Muscle Embedding (MIME) - A Novel Experimental Technique to Facilitate Donor-Cell-Mediated Myogenesis
0 Views •
Cited by 3 •
2017
We describe a novel experimental technique that we call Minimally Invasive Muscle Embedding (MIME), which is based on the evidence that skeletal muscle tissue contains viable myogenic cells that can facilitate donor-cell-mediated myogenesis when implanted into a host muscle.
02:59
Embedding 3D Spheroids in Agarose Gel Drops: A Technique to Preserve Spheroids for Downstream Analysis
0 Views •
2023
This video describes the technique to embed 3D spheroids in agarose gel drops for preservation. The embedded 3D spheroids can be further used for downstream analysis.
07:24
In Situ Hybridization Techniques for Paraffin-Embedded Adult Coral Samples
0 Views •
Cited by 5 •
2018
The goal of this protocol is to perform in situ hybridization on adult coral samples that have been embedded in paraffin and sectioned onto glass slides. This is a qualitative method used to visualize the spatial expression of an RNA anti-sense probe in paraffin-embedded tissues.
FAQs
Resin removal or surface treatment is crucial because embedding can limit access to molecular targets. These steps expose antigenic sites on the section surface, allowing antibodies to bind proteins that might otherwise remain inaccessible. In neural tissue, effective exposure improves the ability to associate a detected marker with a particular neuron, synapse, or subcellular compartment while retaining anatomical context.
Primary antibodies provide target recognition, while labeled secondary antibodies make that recognition visible in the section. After the specimen has been fixed, embedded, and sectioned, the exposed tissue surface can be treated for labeling before microscopy. This arrangement allows researchers to examine proteins such as neurotransmitters, receptors, and synaptic markers without losing their relationship to surrounding neural structures.
Ultrathin sections allow molecular labeling to be examined alongside very fine anatomical detail. This is particularly valuable when a marker must be assigned to a synapse, neuronal process, or other subcellular compartment rather than simply to a broader tissue region. Combined with electron microscopy, the approach supports precise analysis of how molecular components are arranged within neural structures.
Examining neurotransmitters, receptors, synaptic proteins, and other markers in place helps distinguish molecularly defined elements within neural tissue. Their localization can be related to neuronal architecture and subcellular compartments, giving investigators evidence about how synapses are organized and how molecular distributions change during development, injury, or disease. The resulting interpretation links protein identity with neural structure.
A typical workflow begins by fixing the biological specimen, embedding it, and cutting sections, including ultrathin sections when detailed structural analysis is required. Researchers may then remove resin or treat the section surface to expose antigens. Primary antibodies are applied to recognize targets, followed by labeled secondary antibodies, and the sections are examined by light or electron microscopy.
Light microscopy provides a way to examine labeled targets in tissue sections, whereas electron microscopy supports analysis alongside finer ultrastructural context. The appropriate readout depends on whether the investigation emphasizes marker distribution across neural tissue or precise placement within synaptic and other subcellular structures. Both approaches preserve the connection between molecular identity and anatomical organization.
This technique is especially useful when researchers need to connect molecular markers with neural circuitry or synaptic structure. It can support studies of neurotransmitters, receptors, and synaptic proteins, as well as investigations of changes associated with development, injury, or disease. By retaining anatomical relationships during analysis, it helps reveal where molecular alterations occur within the nervous system.