Brain–ring gland complex was dissected out in cold phosphate-buffered saline (PBS) buffer and fixed in 4% paraformaldehyde for 20 min (90 (link)). Brain–ring gland complex was stained overnight at 4°C with chicken anti-GFP (1/500; Life Technologies), and rat anti-Pvr (91 (link)) (1/500; a gift from B. Shilo). Secondary antibodies 488 anti-chicken (1/500) and 647 anti-rat (1/500) were purchased from Invitrogen. For DNA and nuclei staining, we used Hoechst (1/1000) for 10 min at room temperature. We set up the tile scan with the micrometer-thick Z stack on an LSM880 Zeiss microscope with 40× oil objective. Images were acquired with a resolution of 1024 × 1024.
Protocol detail
Brain-Ring Gland Imaging Protocol
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 : Memorial Sloan Kettering Cancer Center
Top 5 similar protocols
Variable analysis
independent variables
- Dissection of brain-ring gland complex
- Staining of brain-ring gland complex with chicken anti-GFP and rat anti-Pvr antibodies
- DNA and nuclei staining with Hoechst
- Fixation of brain-ring gland complex in 4% paraformaldehyde for 20 minutes
- Staining overnight at 4°C
- Staining with secondary antibodies 488 anti-chicken and 647 anti-rat
- Hoechst staining for 10 minutes at room temperature
- Tile scan with micrometer-thick Z stack on LSM880 Zeiss microscope with 40× oil objective
- Image acquisition at a resolution of 1024 × 1024
- 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!