The IVIS Kinetic from Caliper Life Sciences is capable of real time detection and measurement of luminescence and fluorescence data that are generated in vivo (e.g. in mice). The main focus of the research of the applicants is development of nuclei acid transfer technology. A critically important determinant influencing the advancement of this technology to clinical application is availability of vectors that efficiently genes to their target cells. The importance of measuring luciferase activity to determine gene delivery is threefold:
1. It is a convenient surrogate for therapeutic gene expression.
2.The sensitivity is very good, and better than other available methods of measuring gene delivery, and
3. As luciferase activity is linear over a range of at least 7 logs, saturation of the signal only occurs at extremes and quantitation of gene delivery methods can be assessed very accurately.
Specifications of the equipment, which include sensitive quantitative detection of fluorescent labelled molecules (e.g. short interfering RNAs or siRNAs) and reporter gene (e.g. luciferase) activity in vivo, are therefore particularly powerful and useful for advancing therapeutic use of nuclei acids. This capability overcomes the need to perform analysis on sections or extracts of tissue obtained from post mortem. Inclusion of a highly sensitive electron multiplying charge-coupled device (EMCCD) camera for signal enhancement and the ability to reduce exposure times enables the fast kinetic analysis. This means that the IVIS Kinetic can perform quantitative luminescence and fluorescence data detection as a standard imager as well as rapid detector required for kinetic analysis. IVIS systems are supported by a range of practicals accessories. The anaesthetic module, which enables sedation of mice while carrying out analysis, is particularly important for the uses proposed in this application. The IVIS Kinetic also has a light-tight injection port which supports a syringe injector system enabling real time compound and/or substrate administration. A removable pull-out tray is available to facilitate pre-imaging animal procedures. In summary, availability of the equipment allows rapid, sensitive and accurate measurements to be made repeatedly without the need to sacrifice experimental animals. As a result, expenses as well as ethical concerns will be limited by access to the equipment.