Two-photon imaging was performed to observe NET formation in the mouse brain after 24 h of 2.5 mg/kg, intravenous (i.v.) LPS injection. An anesthetized mouse was placed in a customized chamber, and the brain surgery was conducted prior to imaging as previously described (41 (link)42 (link)). NET-defining markers, which are NE Ab Alexa Fluor 488 (Ssc-55549 AF488; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and CitH3 (Cat# ab5103; Abcam), were intravenously inoculated to visualize the NET formation. For the fluorescence imaging of NETs, the CitH3 Ab was conjugated to Alexa Fluor 594 using Ab labeling kits, as per the manufacturer’s instructions (Invitrogen). The blood vessel was visualized by injecting wheat germ agglutinin (WGA) (Cat# 29028; Biotium, Eching, Germany). Acquired Images were analyzed with Volocity, Image J, and Imaris based on previous reports (18 (link)).
Protocol detail
Visualizing NET Formation in Mouse Brain
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.
Corresponding Organization :
Other organizations : Yonsei University
Top 5 similar protocols
Variable analysis
independent variables
- 2.5 mg/kg, intravenous (i.v.) LPS injection
- NET formation in the mouse brain
- Anesthetized mouse
- Brain surgery conducted prior to imaging as previously described (41, 42)
- Positive control: NET-defining markers, including NE Ab Alexa Fluor 488 and CitH3, were intravenously inoculated to visualize the NET formation.
- 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!