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Bioluminescence Imaging Core
Description of the Core Facility:The In-vivo Bioluminescence Molecular Imaging Core began in the Spring of 2002 with the purchase of a Xenogen In-Vivo Imaging System (IVIS). The core is housed in the Laboratory of Molecular Oncology and Cell Cycle Regulation in the Department of Medicine and Abramson Cancer Center, located in the Clinical Research Building at the University of Pennsylvania School of Medicine. The IVIS system provides the capability to perform noninvasive bioluminescence and fluorescence imaging in cell culture and living animals (Figure 1). At the time of its purchase, the unit was one of few at major academic centers and the first in the Philadelphia area.
The current set-up represents a satellite core facility of the greater molecular imaging core at the University of Pennsylvania. Moreover, it is envisioned that the current in-vivo bioluminescence set-up serves as a model for available technology, further technology development as the field of molecular bioluminescence imaging gains more sophistication, e.g. imaging a growing number of reporters and being able to obtain 3-D images and quantitative information about reporter expression. It is anticipated that in the future there will be multiple bioluminescence cores as usage grows, and it is expected that multiple funding sources will support various users or user groups. This serves a purpose to not only provide multiple options during heavy usage, and back-up for inevitable equipment failures or down time, but also the potential to develop and push technology in various directions, e.g. more proteomic imaging or molecular beacon technology versus gene expression imaging. In principle the current set-up can handle all potential applications, but it is clear that in-vivo bioluminescence imaging is a rapidly growing field with numerous applications with high likelihood for widespread need for imaging services. The core is set up to provide service at fee for cost with no profit built in. The costs charged include costs for equipment maintenance, software upgrades, personnel and operational costs. The Xenogen In-vivo Imaging System (IVIS) is a highly sensitive, low light level system optimized for in vivo (whole, living animal) imaging. The system is run by the Living Image software (custom made by Xenogen). The software package automates multiple important aspects of the successful imaging process, including background management, data storage and retrieval, and data quality assurance. Moreover, the installed software package provides guidance for inexperienced users to navigate through complex steps associated with quantitative in vivo imaging. The Living Image “experiment” is simulated and was developed using the Igor data analysis and programming environment. The Living Image experiment file is opened by Igor and creates a custom environment that is used for data acquisition and data analysis. Living Image and Igor run on both Macintosh and Windows computers. Users have unique secure profiles and directories with saved camera settings. The Technical Director provides hands on training for IVIS users interested in bioluminescence or fluorescence imaging of cells in 96- or other-well formats as well as for image acquisition using live animals, data analysis and display, and troubleshooting. The Director and Technical Director are available to assist users with planning of experiments including information about available reporters, sensitivity, background, control experiments, and data interpretation. The Technical Director coordinates scheduling of imaging sessions, and is available during imaging sessions for assistance with questions or difficulties encountered by more experienced users. Users are advised to plan experiments carefully, in consultation with the imaging core in order to not overlook important controls that are needed for complex in-vivo experiments. In addition, users are advised to consult with the core Director and Technical Director in order to conduct reasonable and feasible experiments in terms of the number of samples (plates or animals), cost and effort required, frequency of monitoring in longitudinal studies, and design of multiple reporter experiments. Information is also available to assist users with tissue optics considerations for internal experimental luminescence sources in animals, quantitative measurements, and potential background problems. The Technical Director also provides assistance and advice with data storage, retrieval, as well as import of images generated by other software. Early examples of applications of the currently operational In-vivo Bioluminescence Imaging Core Facility are shown in Figures 2-5.
Bioluminescence Imaging Core (pdf)
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