The eyes were dissected from the rats 24 h after LPS injection and frozen fresh in FSC22 frozen section compound (Leica Microsystems, Wetzlar, Germany) with liquid nitrogen. Dihydroethidium (DHE, Sigma), an oxidative fluorescent dye, was used for immunohistochemical detection of cytosolic superoxide anion (O2−). ROS production was determined by conversion of DHE to ethidium bromide. Square areas of ICB microphotographs (1.44 megapixels in size) were selected, and mean gray values of the DHE signal were evaluated with ImageJ software, as we reported recently [21 (link)]. Two whole ICB areas were evaluated in each eye and then four eyes were examined in each group.
Protocol detail
Quantifying Superoxide Anion in Rat Eyes
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.
Cytosolic
Dihydroethidium
Ethidium bromide
Eyes
Fluorescent dye
Frozen section
Nitrogen
Rats
Superoxide anion
Corresponding Organization :
Other organizations : Hokkaido University, Health Sciences University of Hokkaido
Top 5 similar protocols
Variable analysis
independent variables
- LPS injection
- Cytosolic superoxide anion (O2−) production
- ROS production
- Time of eyes dissection (24 h after LPS injection)
- Freezing method (fresh in FSC22 frozen section compound with liquid nitrogen)
- Use of Dihydroethidium (DHE) for immunohistochemical detection
- Image analysis method (selection of square areas of ICB microphotographs and evaluation of mean gray values of the DHE signal using ImageJ software)
- 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!