Pre-embedding Protocol
Pre-embedding Protocol - Related Videos
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.
11:55
Preparation of Non-human Primate Brain Tissue for Pre-embedding Immunohistochemistry and Electron Microscopy
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Cited by 6 •
2017
Here, we provide an easy, low-cost, and time-efficient protocol to chemically fix primate brain tissue with acrolein fixative, allowing for long-term preservation that is compatible with pre-embedding immunohistochemistry for transmission electron microscopy.
05:15
Investigating Retinal Circuits and Molecular Localization by Pre-Embedding Immunoelectron Microscopy
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2024
This protocol outlines the detailed steps of pre-embedding immunoelectron microscopy, with a focus on exploring synaptic circuits and protein localization in the retina.
02:32
Sectioning, Staining and TEM Imaging of Embedded Exosomes: A Protocol To Visualize the Structural Features of Exosomes Using TEM
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2023
This video describes the technique for preparing a thin section of embedded exosomes. These sections can further be stained and imaged using transmission electron microscopy, or TEM, to study the structural features of exosomes.
02:01
pre-mRNA Processing
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2019
In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA. Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl guanosine). This 5’ cap helps the...
FAQs
Fixation preserves the specimen’s cellular architecture during subsequent processing, helping structures remain available for microscopy. However, reagent exposure must be controlled because the preparation must support both structural preservation and access for later labeling. This balance is especially important when the final analysis combines ultrastructural observation with localization of molecular markers.
Permeabilization helps antibodies or other probes reach relevant sites within the specimen before the supporting medium limits access. Labeling at this stage positions molecular markers for later visualization, while the extent of reagent exposure affects penetration and preservation. The result is more informative localization analysis when structural and molecular information must be interpreted together.
Dehydration and infiltration prepare the processed specimen to enter resin or another embedding matrix. Their inclusion connects earlier preservation and labeling steps with the final physical support needed for microscopy. Because these stages follow reagent exposure and may affect retained structure or marker visibility, careful processing helps maintain reliable downstream ultrastructural or localization results.
The preparation must match both the biological specimen and the intended imaging method. A workflow emphasizing electron microscopy may prioritize preservation of ultrastructural detail, whereas an immunolabeling study also requires molecular markers to remain accessible and detectable. Consequently, fixation, labeling, dehydration, infiltration, and the selected embedding matrix may be adjusted to support the planned analysis.
A typical sequence begins with fixation, followed when needed by permeabilization and labeling with antibodies or other probes. The specimen then undergoes dehydration and infiltration before it is enclosed in resin or another matrix. This ordered workflow first preserves and marks the biological material, then prepares it for physical support during microscopy and targeted analysis.
Important materials include fixatives, permeabilization reagents, antibodies or other probes, dehydration and infiltration reagents, and the resin or alternative matrix used for embedding. Processing conditions also matter, particularly reagent exposure. Controlling these components and conditions helps balance label penetration with structural preservation, improving the reliability of microscopy-based localization and ultrastructural observations.
The approach is particularly useful when researchers need to connect cellular architecture with the location of specific molecular markers. It supports electron microscopy and immunolabeling studies in which preserved ultrastructure and targeted signal must be examined in the same specimen. The resulting preparation can guide analyses of structure, marker position, or both.