Procedure: 1. Turn on the Vevo system using the breaker switch at the back of the system and then the computer standby switch on the left side of the system.
2. Turn on the monitor. Wait for the Windows operating system to start up. The system will automatically log in, and the Vevo 770 software should automatically launch.
3. Select the Measurement package to be used.
4. Select the appropriate RMV scanhead to be used. Fill the nosepiece as follows:
i) Ensure that the acoustic window is securely attached and remove the fill port screw.
ii) Position the RMV scanhead with the fill port pointing upward. Fill the nosepiece with de-ionized water using the supplied water bottle until the nosepiece is almost full. Fill the nosepiece with de-ionized water only
iii) Using the thumb, cover the fill port and tap the scanhead gently several times to dislodge any air bubbles in the nosepiece.
iiii) Check to ensure that the face of the transducer does not contain bubbles. If bubbles appear, tilt the RMV scanhead to dislodge them.
v) Orient the nosepiece so that the remaining air bubbles are placed under the fill port, and finish filling the nosepiece with the syringe.
vi) After removing all the air from the nosepiece, insert and tighten the fill port screw with the screwdriver provided. Be careful to align the fill port screw properly with the threaded hole in the nosepiece. Failure to do so may damage the nosepiece. Do not over-tighten the fill port screw
5. To initialize the desired scanhead, click Initialize. Once a scanhead is initialized, the Study Browser is displayed.
6. Click New and fill out at least the two mandatory study information fields – Operator Name and Study Name.
7. Induce anaesthesia with i.p. hypnorm/hypnovel, or with isofluorane at a dose of 5% in a closed anaesthetizing chamber then maintained at 2.5% by nose cone for the duration of the ultrasound scan.
8. Shave the abdominal fur region of the isofluorane-anaesthetized mouse with an animal clipper first. Completely remove the remaining fur with a standard chemical hair removal cream (Nair) then cleanse the skin thoroughly with water. Do not allow more than 2-3 minutes contact with the animals skin.
9. Optional: Inject up to 4mls sterile PBS i.p. to improve ultrasound transmission in vivo.
10. Apply ultrasound gel liberally over the abdominal region of the mouse.
11. Place the animal on the heated platform of the rail system.
13. Click OK to continue with the study. The Vevo 770 will automatically start acquiring data in B-Mode. Select: Setup > B-Mode Setup, or press to change acquisition parameters. Use the RMV scanhead to image the organ of interest.
14. Press to start and stop data acquisition. The acquired data may be reviewed using the cine loop controls at the bottom of the screen.
15. For 3D-mode, connect the RMV scanhead to the 3D motor stage/rail system. Select Setup > 3D-Mode Setup or press to change the acquisition parameters. Click Init. 3D to initialize the 3D motor stage. Click Start 3D. The 3D motor stage begins stepping along the acquisition route.
16. To store a data loop select Study > Cine Store, or press ‹Cine Store›
17. To store an image select Study > Frame Store, or press ‹Frame Store›
18. To add measurements select Tools > Measurements, or press ‹Measure›
20. To review the study data select Study > Browse Study, or press ‹Browse› Export the study's images and loops from the Study menu option or from the Image Browser.
21. To save the study, Select Study > Close Study, or press ‹Close› to close the study and click Commit Session Data to save the data. To copy the study to an accessible drive, click Copy To.
22. Once the ultrasound examination is concluded, usually within a 15 min period, clean the animal of gel.
23. Switch off the anaesthesia and return the mouse to its cage with food and water ad libitum. Mice normally recover within 3-5 min from isofluorane anesthesia. Mice anaesthetized with injectible agents will require monitoring until recovery (may be up to 4 hours)