Post-embedding Protocol
Post-embedding Protocol - Related Videos
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...
07:46
Data Acquisition Protocol for Determining Embedded Sensitivity Functions
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2016
The data acquisition procedure for determining embedded sensitivity functions is described. Data is acquired and representative results are shown for a residential scale wind turbine blade.
08:47
Epon Post Embedding Correlative Light and Electron Microscopy
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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.
08:46
A Protocol for Rapid Post-mortem Cell Culture of Diffuse Intrinsic Pontine Glioma (DIPG)
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Cited by 55 •
2017
This protocol describes a method for the rapid processing of post-mortem diffuse intrinsic pontine glioma samples for the establishment of patient-derived cell culture models or direct characterization of tumor and microenvironmental cells.
08:08
qPCR Is a Sensitive and Rapid Method for Detection of Cytomegaloviral DNA in Formalin-fixed, Paraffin-embedded Biopsy Tissue
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Cited by 4 •
2014
This protocol describes qPCR detection of cytomegalovirus in formalin-fixed, paraffin-embedded biopsy tissue, which is rapid, sensitive, specific, and useful for interpreting equivocal hematoxylin and eosin or immunohistochemical staining patterns.
FAQs
Control of section thickness, block trimming, and surface preparation determines how consistently the embedded specimen can be examined. Trimming exposes a suitable region of the block, while controlled section thickness helps produce comparable sections. Preparing the section surface supports subsequent staining, labeling, or mounting. Together, these steps improve image quality, reproducibility, and interpretation of biological structures.
The embedding material is handled according to the experimental goal and the properties of the embedded sample. Removing it can make cellular structures or molecular targets accessible for later analysis, whereas retaining it may support the section during examination. This choice affects which structures remain available, how labeling or staining proceeds, and how the final preparation is interpreted.
Post-embedding decisions depend on whether the study emphasizes tissue architecture, cellular or molecular features, or ultrastructural detail. Histological preparations may prioritize staining and labeling that reveal structures across tissue sections, whereas electron microscopy preparations require conditions that preserve and expose ultrastructural features. In both cases, section preparation and mounting must match the intended microscopic examination.
A typical sequence starts with trimming the embedded block to expose the region of interest, followed by cutting sections with controlled thickness. The embedding medium is then either removed or retained, depending on the analysis. Sections may receive stains or labels before being mounted for microscopy. Maintaining consistent surface preparation throughout the sequence supports reliable imaging and comparison.
Stains and labels are applied according to the structures or molecular targets that need to be examined. Their use follows the decision to remove or retain the embedding medium, because that choice affects access to the section. Properly controlled labeling conditions help make cellular structures or targets visible, improving the usefulness of microscopy for biological interpretation.
These protocols prepare embedded specimens for microscopic evaluation of preserved tissue architecture, cellular structures, molecular targets, or ultrastructural features. The resulting sections can support histological or electron microscopy observations, depending on the preparation. Consistent trimming, sectioning, surface preparation, labeling, and mounting make images more reproducible and strengthen interpretation of the observed biological features.