IVIS 200 Series From Revvity

The IVIS 200 series is an in vivo imaging system used for the real-time quantification of bioluminescent/fluorescent markers in vitro and in live animals. The system is built in a light-proof chamber, and includes a 5-position heated stage and a cryogenically cooled CCD. An anesthesia system is hooked to the stage for isoflurane delivery to animals. The CCD is linked to a computer where images can be retrieved and analyzed using the proprietary Living Image® Software.

We have used the IVIS 200 imaging system for over 2 years now. We have used it for the subcutaneous visualization and quantification of bioluminescent tumor cells in nude mice in the presence or absence of chemopreventive agents. Typically, proprietary Caliper Lifesciences breast cancer cell lines overexpressing a luciferase transgene were used. Animals received intraperitoneal injection of luciferin 15 min prior to imaging. Bioluminescence was measured every week, for several weeks following cell implantation.

Prior to any imaging, a short initialization of the machine is needed to reset the stage position and insure that the CCD temperature is optimal. This can take significantly longer if the machine is turned off between users, which is not recommended. After initialization, animals or plates can be laid on the stage for imaging. The 5-position stage allows for the visualization of up to 5 small animals (mice). The grid marker is a nice feature for pre-aligning animals. There is no “locking” safety built in this stage, so it is possible to damage the CCD if the stage is set at this higher point and a wide object or animal is on the stage. Space on the stage even at the lower level can become really limiting since animal limbs cannot be stretched. Using spacers between animals is then mandatory to avoid signal overlap. Although background correction is automatically scheduled when the machine is not in use, it has happened that background correction was not available for a particular imaging setting. The software allows for easy set-up of a wide range of measurement conditions; however, labeling of files and folder is not user-friendly. It is recommended to take particular care of the image attributes for later analysis. The integrated anesthesia system allows for a very good control of gas flow and includes a pumping system to remove excess gas. Still a significant amount of gas can leak inside the chamber. After imaging, all small parts can be separated from the stage and are easy to clean.

The IVIS 200 series performed as advertised. The system allows for easy animal handling and basic maintenance is minimal. In our observations, the collected bioluminescent signals were sustainable and strong, and the proprietary software gives a wide range of flexibility for the quantification of photons flux. Bioluminescent markers should be favored for in vivo imaging with this system.

In conclusion, we were very satisfied with this instrument, which allowed us to obtain the data we sought. A significant period of training and basic experimentation is mandatory to allow for the full use of the system in order to obtain valuable data.