Cells were fixed with half-strength Karnovsky fixative (2% PFA, 2.5% GA, 0.1 M PB, pH 7.4) for 2 h at RT. After scraping, cells were pelleted and embedded in 2% low melting point agarose. The cell pellets were postfixed as previously described [80 (link)] with 1% OsO4, 1.5% K3Fe(CN)6, in 0.065 M Na-cacodylate, stained en bloc with 0.5% uranyl acetate, dehydrated in acetone and embedded in Epon composed of glycid ether (Serva, 21045.02), 2-dodecenylsuccinic acid anhydride (Serva, 20755.02), methyl nadic anhydride (Serva, 29452.03) and benzyldimethylamine (PolyScience, 00141–100) mixed in weight ratio 48:32:20:3. Ultrathin sections were stained with uranyl acetate and lead citrate.
Protocol detail
Ultrastructural Cell Fixation and Embedding
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Acetone
Acid
Agarose
Anhydride
Benzyldimethylamine
Cacodylate
Cell
Citrate
Epon
Ether
Fixative
Pellets
Uranyl acetate
Corresponding Organization : University Medical Center Utrecht
Other organizations : University Medical Center Groningen, University of Groningen, Juntendo University
Top 5 similar protocols
Variable analysis
independent variables
- Fixation with half-strength Karnovsky fixative (2% PFA, 2.5% GA, 0.1 M PB, pH 7.4) for 2 h at RT
- Postfixation with 1% OsO4, 1.5% K3Fe(CN)6, in 0.065 M Na-cacodylate
- En bloc staining with 0.5% uranyl acetate
- Dehydration in acetone
- Embedding in Epon composed of glycid ether, 2-dodecenylsuccinic acid anhydride, methyl nadic anhydride and benzyldimethylamine
- Ultrastructural characteristics of the cells observed in the ultrathin sections
- Cell type (not explicitly mentioned, but assumed to be consistent across the experiment)
- PH of the buffer (0.1 M PB, pH 7.4)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!