We use an Olympus Fluoview1000 two-photon microscope (BX61W1-FV1000, Olympus, Ltd., Tokyo, Japan) with an excitation source of a Spectra-Physics MaiTai HP DeepSee femtosecond Ti:Sa laser. To acquire images (either stacks or single focal planes) from GFP-positive vessels, a long-working-distance (2 mm) water-immersion objective (× 25, NA 1.05) was used for line-scan measurements. The images were taken at 12-bit depth with resolution of 1024×1024 pixels. We chose a scanning rate of 10μs/pixel and 2000 lines in total to follow the velocity of RBCs. The average RBC velocity speed has been reported [8 (link), 15 (link)]. In vivo vessel diameters were measured manually using ImageJ [16 (link)]. RBC velocity and flux were calculated with an automated image-processing algorithm using MATLAB software (The MathWorks Inc., Natick, Massachusetts) [8 (link), 15 (link)].
Protocol detail
Two-Photon Microscopy of RBC Velocity
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.
Top 5 similar protocols
Protocol cited in 1 other protocol
Variable analysis
independent variables
- Excitation source (Spectra-Physics MaiTai HP DeepSee femtosecond Ti:Sa laser)
- Objective (long-working-distance (2 mm) water-immersion objective (× 25, NA 1.05))
- Scanning rate (10μs/pixel)
- Total lines (2000)
- RBC velocity
- RBC flux
- Microscope (Olympus Fluoview1000 two-photon microscope (BX61W1-FV1000))
- Image depth (12-bit)
- Image resolution (1024×1024 pixels)
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!